Synthesis of rocaglamide natural products via photochemical generation of oxidopyrylium species

ABSTRACT

The present invention provides new strategies for the synthesis of compounds of the rocaglamide family and related natural products. In particular, the new biomimetic synthetic approach involves photochemical generation of an oxidopyrylium species from a 3-hydroxychromone derivative followed by 1,3-dipolar cycloaddition of the oxidopyrylium species to a dipolarophile. This approach can be used for the formation of adducts containing an aglain core structure. Methods for the conversion of aglain core structures to aglain, rocaglamide and forbaglin ring systems are also provided. The present invention also relates to the use of rocaglamide/aglain/forbaglin derivatives for the manufacture of medicaments for use in the treatment of cancer or cancerous conditions, disorders associated with cellular hyperproliferation, or NF-κB-dependent conditions.

RELATED APPLICATIONS

This application claims priority to Provisional Application No.60/555,448, filed on Mar. 23, 2004 and entitled “Synthesis of the AglainSkeleton by Photogeneration and Dipolar Cycloaddition of OxidopyryliumsDerived from 3-Hydroxyflavones”, and Provisional Application No.60/612,009 filed on Sep. 22, 2004 and entitled “Synthesis of RocaglamideNatural Products via Photochemical Generation of Oxidopyrylium Species”.Each of these provisional patent applications is incorporated herein byreference in its entirety.

BACKGROUND OF THE INVENTION

The plant genus Aglaia native of the tropical rain forests of Indonesiaand Malaysia is the source of a unique group of densely functionalizednatural products presented on FIG. 1 (P. Proksch et al., Curr. Org.Chem., 2001, 5: 923-938). The rocaglamides, including the parentmolecule (compound 1; M. L. King et al., J. Chem. Soc., Chem. Commun.,1982, 1150-1151) and the recently isolated dioxanyloxy-modifiedderivative silvestrol (compound 2; B. Y. Hwang et al., J. Org. Chem.,2004, 69: 3350-3358), possess a cyclopenta[b]tetrahydrobenzofuran ringsystem (presented in red on FIG. 1). The structurally related aglains(e.g., compounds 3 and 4), which contain a cyclopenta[bc]benzopyranstructure (presented in blue on FIG. 1), have also been isolated fromAglaia (V. Dumontet et al., Tetrahedron, 1996, 52: 6931-6942). Theforbaglins (e.g., compound 5) are benzo[b]oxepines (in green on FIG. 1)derived from formal oxidative cleavage of the aglain core.

The rocaglamides have been shown to exhibit potent anticancer (M. L.King et al., J. Chem. Soc., Chem. Commun., 1982, 1150-1151) andantileukemic activity (S. K. Lee et al., Chem. Biol. Interact., 1998,115: 215-228), as well as NF-κB inhibitory activity at nanomolarconcentrations in human T cells (B. Baumann et al., J. Biol. Chem.,2002, 277: 44791-44800). The rocaglate silvestrol 2 displays cytotoxicactivity against human cancer cells comparable to the anticancer drugTaxol (B. Y. Hwang et al., J. Org. Chem., 2004, 69: 3350-3358).

As proposed by Proksch (P. Proksch et al., Curr. Org. Chem., 2001, 5:923-938) and Bacher (M. Bacher et al., Phytochemistry, 1999, 52:253-263), and as shown on FIG. 2, the rocaglamides may bebiosynthetically derived from reaction of trimethoxy-substituted3-hydroxyflavone with cinnamide derivatives to afford the aglain corefollowed by skeletal rearrangement.

Although the rocaglamides have been the subject of a number of syntheticinvestigations (see, for example, G. A. Kraus and J. O. Sy, J. Org.Chem., 1989, 54: 77-83; B. Trost et al., J. Am. Chem. Soc., 1990, 112:9022-9024), including a biomimetic approach involving a [2+2]photocycloaddition (H. C. Hailes et al., Tetrahedron Lett., 1993, 34:5313-5316), syntheses of the related aglain (V. Dumontet et al.,Tetrahedron, 1996, 52: 6931-6942), aglaforbesin (V. Dumontet et al.,Tetrahedron, 1996, 52: 6931-6942), or forbaglins have not been reported.Moreover, a unified synthetic approach to these molecules based onbiosynthetic considerations still remains to be developed.

SUMMARY OF THE INVENTION

The present invention provides new methods for the synthesis of naturalproducts. In particular, the invention encompasses novel strategies forthe biomimetic preparation of compounds in therocaglamide/aglain/forbaglin family.

More specifically, one aspect of the present invention relates to theuse of a photochemically generated oxidopyrylium species as anintermediate in a chemical reaction. In certain preferred embodiments,the photochemical reaction leading to the formation of the oxidopyryliumspecies comprises an excited state intramolecular proton transfer.

For example, the oxidopyrylium species may be produced by photochemicalirradiation of a 3-hydroxychromone derivative (I) with the followingchemical structure:

wherein R₁, R₂, R₃, R₄ and R are identical or different and selectedfrom the group consisting of hydrogen, halogen, hydroxy, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.

In particular, the oxidopyrylium species may be produced byphotochemical irradiation of a 3-hydroxyflavone derivative (II) with thefollowing chemical structure:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are identical or differentand selected from the group consisting of hydrogen, halogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl. In certainpreferred embodiments, the 3-hydroxyflavone derivative has one of thefollowing chemical structures:

Alternatively, the oxidopyrylium species may be produced byphotochemical irradiation of a 5-hydroxy-2,3-dihydro-pyran-4-onederivative (III) with the following chemical structure:

wherein R₁, R₂, R₃, R₄ and R are identical or different and selectedfrom the group consisting of hydrogen, halogen, hydroxy, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.

In certain embodiments, the photochemically generated oxidopyryliumspecies is used as an intermediate in a cycloaddition, for example a1,3-dipolar cycloaddition, leading to the formation of an adduct.

Another aspect of the present invention relates to a method comprisingsteps of: photochemically generating an oxidopyrylium species; andreacting the oxidopyrylium species thus obtained with a dipolarophile.In certain preferred embodiments, the oxidopyrylium species is producedby photoinduced excited state intramolecular proton transfer of a3-hydroxychromone derivative of chemical structure (I), or a3-hydroxyflavone derivative of chemical structure (II) or a5-hydroxy-2,3-dihydro-pyran-4-one derivative of chemical structure(III), as described above.

In certain embodiments, the reaction between the oxidopyrylium speciesand the dipolarophile (e.g., a cinnamate derivative) comprises acycloaddition, (e.g., a 1,3-dipolar cycloaddition), and results in theformation of an adduct. Preferably, the adduct comprises an aglain corestructure. In other embodiments, the inventive method further comprisesconverting the adduct formed. For example, when the adduct formedcomprises an aglain core structure, converting the adduct may result inthe formation of a ring system selected from the group consisting of anaglain ring system, a rocaglamide ring system, and a forbaglin ringsystem.

In another aspect, the present invention provides a method for preparinga compound containing an aglain core structure, said method comprisingsteps of: producing an oxidopyrylium species (I_(T)) by photoinducedexcited state intramolecular proton transfer of a 3-hydroxychromonederivative (I); and reacting the oxidopyrylium species with adipolarophile (IV) to obtain the aglain core-containing compound (V).Compounds (I), (I_(T)), (IV), and (V) have the following chemicalstructures:

wherein R₁, R₂, R₃, R₄, R, R_(a) and R_(b) are identical or differentand selected from the group consisting of hydrogen, halogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.

Alternatively, the method for preparing a compound containing an aglaincore structure may comprising steps of: producing an oxidopyryliumspecies (II_(T)) by photoinduced excited state intramolecular protontransfer of a 3-hydroxyflavone derivative (II); and reacting theoxidopyrylium species with a dipolarophile (IV) to obtain the aglaincore-containing compound (V′). Compounds (II), (II_(T)), (IV), and (V′)have the following chemical structures:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein each occurrence of R_(x) is independentlyselected from the group consisting of hydrogen, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, andheteroaryl.

In certain preferred embodiments of these methods, the dipolarophile(IV) is a cinnamate derivative with the following chemical structure:

wherein R¹ is selected from the group consisting of hydrogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic,alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic,aryl, heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, and aprotecting group; andwherein R², R³, R⁴, R⁵, and R⁶ are identical or different and selectedfrom the group consisting of hydrogen, halogen, hydroxy, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.

In certain embodiments, the inventive methods further compriseconverting the compound with an aglain core structure obtained. Forexample, the aglain core-containing compound may be converted into acompound with a ring system selected from the group consisting of anaglain ring system, a rocaglamide ring system, and a forbaglin ringsystem. Conversion into a compound with an aglain ring system mayinvolve a reduction. Conversion into a compound with a rocaglamide ringsystem may comprise an α-ketol (acyloin) rearrangement (preferably underbasic conditions), and optionally a hydroxyl-directed reduction.Conversion into a compound with a forbaglin ring system may comprise anoxidative cleavage.

In another aspect, the present invention relates to a method forpreparing an aglain derivative, the method comprising steps of:producing an oxidopyrylium species (I_(T)) by photoinduced excited stateintramolecular proton transfer of a 3-hydroxychromone derivative (I);reacting the oxidopyrylium species with a dipolarophile (IV) to obtain acompound with an aglain core structure (V); and converting the compoundwith the aglain core structure into an aglain derivative (VI). Compounds(I), (I_(T)), (IV), and (V) are as described above and compound (VI) hasthe following chemical structure:

wherein R₁, R₂, R₃, R₄, R, R_(a) and R_(b) are identical or differentand selected from the group consisting of hydrogen, halogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl; andwherein R′ is selected from the group consisting of hydrogen, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —S(O)R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, and —N(R_(x))S(O)₂R_(x), wherein eachoccurrence of R_(x) is independently selected from the group consistingof hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, and heteroaryl.

Alternatively, the method for preparing an aglain derivative comprisessteps of: producing an oxidopyrylium species (II_(T)) by photoinducedexcited state intramolecular proton transfer of a 3-hydroxyflavonederivative (II); reacting the oxidopyrylium species with a dipolarophile(IV) to obtain a compound with an aglain core structure (V′); andconverting the compound with an aglain core structure into an aglainderivative (VI′). Compounds (II), (II_(T)), (IV), and (V′) are asdescribed above and compound (VI′) has the following chemical structure:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein each occurrence of R_(x) is independentlyselected from the group consisting of hydrogen, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, andheteroaryl; andwherein R′ is selected from the group consisting of hydrogen, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —S(O)R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, and —N(R_(x))S(O)₂R_(x), wherein eachoccurrence of R_(x) is independently selected from the group consistingof hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, and heteroaryl.

In certain embodiments of these methods, the dipolarophile (IV) is acinnamate derivative as described above.

In certain preferred embodiments, converting the compound with an aglaincore structure into an aglain derivative involves a reduction, forexample carried out in the presence of NaBH₄, Me₄NBH(OAc)₃ or anothersuitable reducing agent. Alternatively, addition of nucleophiles, e.g.,Grignard or alkyllithium reagents, may be performed to convert theaglain core-containing compound into an aglain derivative.

In another aspect, the present invention relates to a method forpreparing a rocaglamide derivative, the method comprising steps of:producing an oxidopyrylium species (I_(T)) by photoinduced excited stateintramolecular proton transfer of a 3 hydroxychromone derivative (I);reacting the oxidopyrylium species obtained with a dipolarophile (IV) toobtain a compound with an aglain core structure (V); and converting thecompound with an aglain core structure into a rocaglamide derivative(VII). Compounds (I), (I_(T)), (IV), and (V) are as described above and(VII) has the following chemical structures:

wherein R₁, R₂, R₃, R₄, R, R_(a) and R_(b) are identical or differentand selected from the group consisting of hydrogen, halogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.

Alternatively, the method for preparing a rocaglamide derivativecomprises steps of: producing an oxidopyrylium species (II_(T)) byphotoinduced excited state intramolecular proton transfer of a3-hydroxyflavone derivative (II); reacting the oxidopyrylium speciesobtained with a dipolarophile (IV) to obtain a compound with an aglaincore structure (V′); and converting the compound with an aglain corestructure into a rocaglamide derivative (VII′). Compounds (II),(II_(T)), (IV), and (V′) are as described above and (VII′) has thefollowing chemical structures:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein each occurrence of R_(x) is independentlyselected from the group consisting of hydrogen, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, andheteroaryl.

In certain embodiments of these methods, the dipolarophile (IV) is acinnamate derivative as described above.

In certain preferred embodiments, converting the compound with an aglaincore structure into a rocaglamide derivative comprises an α-ketol(acyloin) rearrangement and optionally a hydroxyl-directed reduction.Preferably, the α-ketol rearrangement is carried out under basicconditions.

Another aspect of the present invention relates to a method forpreparing a rocaglamide derivative, the method comprising steps of:producing an oxidopyrylium species (I_(T)) by photoinduced excited stateintramolecular proton transfer of a 3-hydroxychromone derivative (I);reacting the oxidopyrylium species obtained with a dipolarophile (IV) toobtain a compound with an aglain core structure (V); and converting thecompound with an aglain core structure into a rocaglamide derivative(VIII). Compounds (I), (I_(T)), (IV), and (V) are as described andcompound (VIII) has the following chemical structures:

wherein R₁, R₂, R₃, R₄, R, R_(a) and R_(b) are identical or differentand selected from the group consisting of hydrogen, halogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl; andwherein R′ is selected from the group consisting of hydrogen, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —S(O)R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, and —N(R_(x))S(O)₂R_(x), wherein eachoccurrence of R_(x) is independently selected from the group consistingof hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, and heteroaryl.

Alternatively, the method for preparing a rocaglamide derivativecomprises steps of: producing an oxidopyrylium species (II_(T)) byphotoinduced excited state intramolecular proton transfer of a3-hydroxyflavone derivative (II); reacting the oxidopyrylium speciesobtained with a dipolarophile (IV) to obtain a compound with an aglaincore structure (V′); and converting the compound with an aglain corestructure into a rocaglamide derivative (VIII′). Compounds (II),(II_(T)), (IV), and (V′) are as described above and compound (VIII′) hasthe following chemical structures:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein each occurrence of R_(x) is independentlyselected from the group consisting of hydrogen, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, andheteroaryl; andwherein R′ is selected from the group consisting of hydrogen, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —S(O)R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, and —N(R_(x))S(O)₂R_(x), wherein eachoccurrence of R_(x) is independently selected from the group consistingof hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, and heteroaryl.

In certain embodiments of these methods, the dipolarophile (IV) is acinnamate derivative as described above.

In certain preferred embodiments, converting the compound with an aglaincore structure into a rocaglamide derivative comprises an α-ketol(acyloin) rearrangement and optionally a hydroxyl-directed reduction.Preferably, the α-ketol rearrangement is carried out under basicconditions.

In another aspect, the present invention relates to a method forpreparing a forbaglin derivative, the method comprising steps of:producing an oxidopyrylium species (I_(T)) by photoinduced excited stateintramolecular proton transfer of a 3-hydroxychromone derivative (I);reacting the oxidopyrylium species obtained with a dipolarophile (IV) toobtain a compound with an aglain core structure (V); and converting thecompound with an aglain core into a forbaglin derivative (IX). Compounds(I), (I_(T)), (IV), and (V) are as described above and compound (IX) hasthe following chemical structures:

wherein R₁, R₂, R₃, R₄, R, R″, R_(a) and R_(b) are identical ordifferent and selected from the group consisting of hydrogen, halogen,hydroxy, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl,thioaryl, acyl, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl,arylamino, amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃,—CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.

Alternatively, the method for preparing a forbaglin derivative comprisessteps of: producing an oxidopyrylium species (II_(T)) by photoinducedexcited state intramolecular proton transfer of a 3-hydroxyflavonederivative (II); reacting the oxidopyrylium species obtained with adipolarophile (IV) to obtain a compound with an aglain core structure(V′); and converting the compound with an aglain core into a forbaglinderivative (IX′). Compounds (II), (II_(T)), (IV), and (V′) are asdescribed above and compound (IX′) has the following chemicalstructures:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R″, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein each occurrence of R_(x) is independentlyselected from the group consisting of hydrogen, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, andheteroaryl.

In certain embodiments of these methods, the dipolarophile (IV) is acinnamate derivative as described above.

In certain preferred embodiments, converting the compound with an aglaincore structure into a forbaglin derivative comprises an oxidativecleavage, for example, an oxidative cleavage carried out in the presenceof Pb(OAc)₄.

Another aspect of the present invention relates to aglain corecontaining compounds (V) and (V′), aglain derivatives (VI) and (VI′),rocaglamide derivatives (VII), (VII′), (VIII) and (VIII′), and forbaglinderivatives (IX) and (IX′) prepared by the methods disclosed herein.

Another aspect of the present invention relates to the use of thesecompounds and derivatives for the manufacture of medicaments for use inthe treatment of disease states including cancer or cancerousconditions, conditions associated with cellular hyperproliferation, andNF-κB-associated conditions.

For example, cancer and cancerous conditions that may be treated by suchmedicaments include leukemia, sarcoma, breast, colon, bladder,pancreatic, endometrial, head and neck, mesothelioma, myeloma,oesophagal/oral, testicular, thyroid, cervical, bone, renal, uterine,prostate, brain, lung, ovarian, skin, liver and bowel and stomachcancers, tumors and melanomas. Conditions associated with cellularhyperproliferation that can be treated using the inventive medicamentsmay be selected from the group consisting of atherosclerosis,restenosis, rheumatoid arthritis, osteoarthritis, inflammatoryarthritis, psoriasis, periodontal disease and virally induced cellularhyperproliferation. NF-κB associated conditions that can be treatedusing the medicaments disclosed herein may be selected from the groupconsisting of immunological disorders, septic shock, transplantrejection, radiation damage, reperfusion injuries after ischemia,arteriosclerosis and neurodegenerative diseases.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 shows the chemical structures of Rocaglamide and related naturalcompounds isolated from the plant genus Aglaia.

FIG. 2 shows a reaction scheme proposed by Proksch and coworkers (Curr.Org. Chem., 2001, 5: 923-938) for the biosynthetic preparation of therocaglamides.

FIG. 3 shows an embodiment of the inventive unified biomimetic approachto the synthesis of Aglains-Forbaglins-Rocaglamides.

FIG. 4 is a scheme showing the excited state intramolecular protontransfer (ESIPT) process and fluorescence emission taking place uponphotoirradiation of the parent molecule, 3-hydroxyflavone.

FIG. 5 shows the reaction of photochemical [3+2] cycloaddition between3-hydroxyflavone 13 and methyl cinnamate 14.

FIG. 6 shows the ¹H-NMR (400 MHz, CDCl₃) (A) and ¹³C-NMR (75 MHz, CDCl₃)(B) spectra recorded for compound 16, which results from photochemical[3+2] cycloaddition between 3-hydroxyflavone 13 and methyl cinnamate 14.

FIG. 7 shows the ¹H-NMR spectrum (400 MHz, CD₃CN) of a mixture of3-hydroxyflavone 13 (1 equivalent) and methyl cinnamate 14 (5equivalents) after 2 hours of irradiation. The chemical structure ofmethyl cinnamate 14 is presented in red and the chemical structure ofcompound 16, the main product of the reaction, is presented in blue.

FIG. 8 shows parts (3 to 5 ppm) of expanded ¹H-NMR spectra (400 MHz,CD₃CN) recorded for compound 16 (FIG. 8(A)); and for a mixture of 3hydroxyflavone 13 and methyl cinnamate 14 after 2 hours of irradiation(FIG. 8(B)).

FIG. 9 shows an example of chemical conversion of an aglain corestructure to forbaglin and rocaglamide ring systems.

FIG. 10 shows the ¹H-NMR (400 MHz, CDCl₃) (A) and ¹³C-NMR (75 MHz,CDCl₃) (B) spectra recorded for compound 23.

FIG. 11 is a scheme presenting an example of synthesis of (±) methylrocaglate from trimethoxy-substituted 3-hydroxyflavone.

FIG. 12 shows the reaction sequence used to synthesizetrimethoxy-substituted 3-hydroxyflavone 24.

FIG. 13 shows the chemical structures of compound 27, keto isomer 27′and enol isomer 27″.

FIG. 14 shows the ¹H-NMR (400 MHz, CDCl₃) (A) and ¹³C-NMR (75 MHz,CDCl₃) (B) spectra recorded for compound 28.

FIG. 15 shows the ¹H-NMR (400 MHz, CDCl₃) (A) and ¹³C-NMR (75 MHz,CDCl₃) (B) spectra recorded for compound 29.

FIG. 16 shows the HMQC spectrum of synthetic exo methyl rocaglate 29(500 MHz, CHCl₃, 25° C.).

FIG. 17 shows the HMBC spectrum of synthetic exo methyl rocaglate 29(500 MHz, CHCl₃, 25° C.).

FIG. 18 shows the HMBC spectrum of synthetic exo methyl rocaglate 29(500 MHz, CHCl₃, 25° C.).

FIG. 19 shows the chemical structures of compounds 30 and 31, obtainedfrom chemical modifications of compounds 16 and 15, respectively.

FIG. 20 shows the X-ray Crystal Structure of Compound 30.

FIG. 21 shows the X-ray Crystal Structure of Compound 31.

DEFINITIONS

Throughout the specification, several terms are employed that aredefined in the following paragraphs.

The terms “oxidopyrylium species” and “oxidopyrylium ylide species” areused herein interchangeably. An oxidopyrylium species is a dipolarentity, i.e., an electrically neutral molecule carrying a positivecharge and a negative charge in one of its major canonical descriptions.In the context of the present invention, an oxidopyrylium speciespreferably comprises the following chemical group/motif:

Preferred oxidopyrylium species have chemical structure (I_(T)) or(II_(T)). In most of the inventive methods provided herein, anoxidopyrylium species is photochemically generated and used as anintermediate in a chemical reaction.

The terms “photochemically generated” and “generated in a photochemicalreaction” are used herein interchangeably to characterize a chemicalentity whose formation is caused/initiated by absorption of ultraviolet,visible, or infrared radiation. Similarly, a chemical process orreaction is “photoinduced” if it is caused/initiated by absorption ofultraviolet, visible, or infrared radiation. A wide variety of chemicalprocesses/reactions may be photoinduced including, but not limited to,additions, cyclizations, eliminations, enolizations, rearrangements,isomerizations, oxidations, reductions, substitutions, and the like.

As used herein, the term “intermediate” refers to a molecular entitywith a lifetime appreciably longer than a molecular vibration that isformed (directly or indirectly) from one or more reactants and reactsfurther to give (either directly or indirectly) the product(s) of achemical reaction.

The term “cycloaddition”, as used herein, refers to a chemical reactionin which two or more π-electron systems (e.g., unsaturated molecules orparts of the same unsaturated molecule) combine to form a cyclic productin which there is a net reduction of the bond multiplicity. In acycloaddition, the π electrons are used to form new σ bonds. The productof a cycloaddition is called an “adduct” or a “cycloadduct”. Differenttypes of cycloaddition are known in the art including, but not limitedto, 1,3-dipolar cycloadditions and Diels-Alder reactions.

As used herein, the term “converting” refers to a process or reactionthat is aimed at modifying a chemical compound. A variety of processesor reactions can be used to convert or modify a chemical compoundincluding, but not limited to, additions, eliminations, substitutions,oxidations, reductions, enolizations, rearrangements, isomerizations,and the like.

The term “aliphatic”, as used herein, includes both saturated andunsaturated, straight chain (i.e., unbranched) or branched aliphatichydrocarbons, which are optionally substituted with one or morefunctional groups. As will be appreciated by one of ordinary skill inthe art, the term “aliphatic” is intended herein to include, but is notlimited to, alkyl, alkenyl, or alkynyl moieties. As used herein, theterm “alkyl” includes straight and branched alkyl groups. An analogousconvention applies to other generic terms such as “alkenyl”, “alkynyl”and the like. Furthermore, as used herein, the terms “alkyl”, “alkenyl”,“alkynyl” and the like encompass both substituted and unsubstitutedgroups. In certain embodiments, as used herein, “lower alkyl” is used toindicate those alkyl groups (substituted, unsubstituted, branched orunbranched) having 1-6 carbon atoms. “Lower alkenyl” and “lower alkynyl”respectively include corresponding 1-6 carbon moieties.

In certain embodiments, the alkyl, alkenyl and alkynyl groups employedin the invention contain 1-20; 2-20; 3-20; 4-20; 5-20; 6-20; 7-20 or8-20 aliphatic carbon atoms. In certain other embodiments, the alkyl,alkenyl, and alkynyl groups employed in the invention contain 1-10;2-10; 3-10; 4-10; 5-10; 6-10; 7-10 or 8-10 aliphatic carbon atoms. Inyet other embodiments, the alkyl, alkenyl, and alkynyl groups employedin the invention contain 1-8; 2-8; 3-8; 4-8; 5-8; 6-20 or 7-8 aliphaticcarbon atoms. In still other embodiments, the alkyl, alkenyl, andalkynyl groups employed in the invention contain 1-6; 2-6; 3-6; 4-6 or5-6 aliphatic carbon atoms. In yet other embodiments, the alkyl,alkenyl, and alkynyl groups employed in the invention contain 1-4; 2-4or 3-4 carbon atoms. Illustrative aliphatic groups thus include, but arenot limited to, for example, methyl, ethyl, n-propyl, isopropyl, allyl,n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, sec-pentyl,isopentyl, tert-pentyl, n hexyl, sec-hexyl, moieties and the like, whichagain, may bear one or more substituents. Alkenyl groups include, butare not limited to, for example, ethenyl, propenyl, butenyl,1-methyl-2-buten-1-yl, and the like. Representative alkynyl groupsinclude, but are not limited to, ethynyl, 2-propynyl(propargyl),1-propynyl and the like.

The term “alicyclic”, as used herein, refers to compounds which combinethe properties of aliphatic and cyclic compounds and include, but arenot limited to, monocyclic, or polycyclic aliphatic hydrocarbons andbridged cycloalkyl compounds, which are optionally substituted with oneor more functional groups. As will be appreciated by one of ordinaryskill in the art, the term “alicyclic” is intended herein to include,but is not limited to, cycloalkyl, cycloalkenyl, and cycloalkynylmoieties, which are optionally substituted with one or more functionalgroups. Illustrative alicyclic groups thus include, but are not limitedto, for example, cyclopropyl, —CH₂-cyclopropyl, cyclobutyl,—CH₂-cyclobutyl, cyclopentyl, —CH₂-cyclopentyl, cyclohexyl,—CH₂-cyclohexyl, cyclohexenylethyl, cyclohexanylethyl, norbornylmoieties and the like, which again, may bear one or more substituents.

The term “alkoxy” or “alkyloxy”, as used herein refers to a saturated(i.e., O-alkyl) or unsaturated (i.e., O-alkenyl and O-alkynyl) groupattached to the parent molecular moiety through an oxygen atom. Incertain embodiments, the alkyl group contains 1-20; 2-20; 3-20; 4-20;5-20; 6-20; 7-20 or 8-20 aliphatic carbon atoms. In certain otherembodiments, the alkyl group contains 1-10; 2-10; 3-10; 4-10; 5-10;6-10; 7-10 or 8-10 aliphatic carbon atoms. In yet other embodiments, thealkyl, alkenyl, and alkynyl groups employed in the invention contain1-8; 2-8; 3-8; 4-8; 5-8; 6-20 or 7-8 aliphatic carbon atoms. In stillother embodiments, the alkyl group contains 1-6; 2-6; 3-6; 4-6 or 5-6aliphatic carbon atoms. In yet other embodiments, the alkyl groupcontains 1-4; 2-4 or 3-4 aliphatic carbon atoms. Examples of alkoxygroups, include but are not limited to, methoxy, ethoxy, propoxy,isopropoxy, n-butoxy, i-butoxy, sec-butoxy, tert-butoxy, neopentoxy,n-hexoxy and the like.

The term “thioalkyl”, as used herein, refers to a saturated (i.e.,S-alkyl) or unsaturated (i.e., S-alkenyl and S-alkynyl) group attachedto the parent molecular moiety through a sulfur atom. In certainembodiments, the alkyl group contains 1-20 aliphatic carbon atoms. Incertain other embodiments, the alkyl group contains 1-10 aliphaticcarbon atoms. In yet other embodiments, the alkyl, alkenyl, and alkynylgroups employed in the invention contain 1-8 aliphatic carbon atoms. Instill other embodiments, the alkyl group contains 1-6 aliphatic carbonatoms. In yet other embodiments, the alkyl group contains 1-4 aliphaticcarbon atoms. Examples of thioalkyl groups include, but are not limitedto, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, andthe like.

The term “alkylamino” refers to a group having the structure —NHR_(a)wherein R_(a) is aliphatic or alicyclic, as defined herein. The term“amino alkyl” refers to a group having the structure NH₂R_(a)—, whereinR_(a) is aliphatic or alicyclic, as defined herein. In certainembodiments, the aliphatic or alicyclic group contains 1-20 aliphaticcarbon atoms. In certain other embodiments, the aliphatic or alicyclicgroup contains 1-10 aliphatic carbon atoms. In still other embodiments,the aliphatic or alicyclic group contains 1-6 aliphatic carbon atoms. Inyet other embodiments, the aliphatic or alicyclic group contains 1-4aliphatic carbon atoms. In yet other embodiments, R_(a) is an alkyl,alkenyl, or alkynyl group containing 1-8 aliphatic carbon atoms.Examples of alkylamino groups include, but are not limited to,methylamino, ethylamino, iso-propylamino and the like.

Some examples of substituents (or functional groups) of theabove-described aliphatic (and other) moieties of compounds of theinvention include, but are not limited to aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylaryl, heteroalkylaryl, alkylheteroaryl,heteroalkylheteroaryl, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy,alkylthio, arylthio, heteroalkylthio, heteroarylthio, F, Cl, Br, I, —OH,—NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂NH₂,—CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)R_(x),—OCO₂R_(x), —OC(═O)N(R_(x))₂, —N(R_(x))₂, —OR_(x), —SR_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x), —N(R_(x))S(O)₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —S(O)₂N(R_(x))₂, wherein each occurrence ofR_(x) independently includes, but is not limited to, aliphatic,alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl,alkylheteroaryl, heteroalkylaryl or heteroalkylheteroaryl, wherein anyof the aliphatic, alicyclic, heteroaliphatic, heterocyclic, alkylaryl,or alkylheteroaryl groups described above and herein may be substitutedor unsubstituted, branched or unbranched, saturated or unsaturated, andwherein any of the aryl or heteroaryl substituents described above andherein may be substituted or unsubstituted.

In general, the term “aromatic moiety” or “aromatic”, as used herein,refers to a stable mono- or poly-cyclic, unsaturated moiety havingpreferably 3-14 carbon atoms, each of which may be substituted orunsubstituted. In certain embodiments, the term “aromatic moiety” refersto a planar ring having p-orbitals perpendicular to the plane of thering at each ring atom and satisfying the Huckel rule where the numberof π electrons in the ring is (4n+2) wherein n is an integer. A mono- orpolycyclic, unsaturated moiety that does not satisfy one or all of thesecriteria for aromaticity is defined herein as “non-aromatic”, and isencompassed by the term “alicyclic”.

In general, the term “heteroaromatic”, as used herein, refers to astable mono- or polycyclic, unsaturated moiety having preferably 3-14carbon atoms, each of which may be substituted or unsubstituted; andcomprising at least one heteroatom selected from O, S and N within thering (i.e., in place of a ring carbon atom). In certain embodiments, theterm “heteroaromatic moiety” refers to a planar ring comprising at leastone heteroatom, having p-orbitals perpendicular to the plane of the ringat each ring atom, and satisfying the Huckel rule where the number of πelectrons in the ring is (4n+2) wherein n is an integer.

It will also be appreciated that aromatic and heteroaromatic moieties,as defined herein may be attached via an alkyl or heteroalkyl moiety andthus also include -(alkyl)aromatic, -(heteroalkyl)aromatic,-(heteroalkyl)heteroaromatic, and -(heteroalkyl)heteroaromatic moieties.Thus, as used herein, the phrases “aromatic or heteroaromatic moieties”and “aromatic, heteroaromatic, -(alkyl)aromatic, -(heteroalkyl)aromatic,-(heteroalkyl)heteroaromatic, and -(heteroalkyl)heteroaromatic” areinterchangeable. Substituents include, but are not limited to, any ofthe previously mentioned substituents, i.e., the substituents recitedfor aliphatic moieties, or for other moieties as disclosed herein,resulting in the formation of a stable compound.

The term “aryl”, as used herein, does not differ significantly from thecommon meaning of the term in the art, and refers to an unsaturatedcyclic moiety comprising at least one aromatic ring. In certainembodiments, the term“aryl” refers to a mono- or bicyclic carbocyclicring system having one or two aromatic rings including, but not limitedto, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like.

The term “heteroaryl”, as used herein, refers to a cyclic aromaticradical having from five to ten ring atoms of which one ring atom isselected from S, O and N; zero, one or two ring atoms are additionalheteroatoms independently selected from S, O and N; and the remainingring atoms are carbon, the radical being joined to the rest of themolecule via any of the ring atoms, such as, for example, pyridyl,pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, and the like.

It will be appreciated that aryl and heteroaryl groups (includingbicyclic aryl groups) can be unsubstituted or substituted, whereinsubstitution includes replacement of one or more of the hydrogen atomsthereon independently with any one or more substituents. Suitablesubstituents include, but are not limited to, any of the previouslymentioned substituents, i.e., the substituents recited for aliphaticmoieties, or for other moieties as disclosed herein, resulting in theformation of a stable compound.

The term “cycloalkyl”, as used herein, refers specifically to groupshaving three to seven, preferably three to ten carbon atoms. Suitablecycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl and the like, which, as in the caseof aliphatic, alicyclic, heteroaliphatic or heterocyclic moieties, mayoptionally be substituted with any of the previously mentionedsubstituents.

The term “heteroaliphatic”, as used herein, refers to aliphatic moietiesin which one or more carbon atoms in the main chain have beensubstituted with a heteroatom. Thus, a heteroaliphatic group refers toan aliphatic chain which contains one or more oxygen, sulfur, nitrogen,phosphorus or silicon atoms, e.g., in place of carbon atoms.Heteroaliphatic moieties may be linear or branched, and saturated orunsaturated. In certain embodiments, heteroaliphatic moieties aresubstituted by independent replacement of one or more of the hydrogenatoms thereon with one or more of the previously mentioned substituents.

The term “heterocycloalkyl”, “heterocycle” or “heterocyclic”, as usedherein, refers to compounds which combine the properties ofheteroaliphatic and cyclic compounds and include, but are not limitedto, saturated and unsaturated mono- or polycyclic cyclic ring systemshaving 5-16 atoms wherein at least one ring atom is a heteroatomselected from O, S and N (wherein the nitrogen and sulfur heteroatomsmay optionally be oxidized), wherein the ring systems are optionallysubstituted with one or more functional groups, as defined herein. Incertain embodiments, the term “heterocycloalkyl”, “heterocycle” or“heterocyclic” refers to a non-aromatic 5-, 6- or 7-membered ring or apolycyclic group wherein at least one ring atom is a heteroatom selectedfrom O, S and N (wherein the nitrogen and sulfur heteroatoms mayoptionally be oxidized), including, but not limited to, a bi- ortri-cyclic group, comprising fused six-membered rings having between oneand three heteroatoms independently selected from oxygen, sulfur andnitrogen, wherein (i) each 5-membered ring has 0 to 2 double bonds, each6-membered ring has 0 to 2 double bonds and each 7-membered ring has 0to 3 double bonds, (ii) the nitrogen and sulfur heteroatoms mayoptionally be oxidized, (iii) the nitrogen heteroatom may optionally bequaternized, and (iv) any of the above heterocyclic rings may be fusedto an aryl or heteroaryl ring. Representative heterocycles include, butare not limited to, heterocycles such as furanyl, thiofuranyl, pyranyl,pyrrolyl, pyrazolyl, imidazolyl, thienyl, pyrrolidinyl, pyrazolinyl,pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl,oxazolyl, oxazolidinyl, isooxazolyl, isoxazolidinyl, dioxazolyl,thiadiazolyl, oxadiazolyl, tetrazolyl, triazolyl, thiatriazolyl,oxatriazolyl, thiadiazolyl, oxadiazolyl, morpholinyl, thiazolyl,thiazolidinyl, isothiazolyl, isothiazolidinyl, dithiazolyl,dithiazolidinyl, tetrahydrofuryl, and benzofused derivatives thereof.The term “heterocycle, or heterocycloalkyl or heterocyclic” alsoencompasses heterocycle, or heterocycloalkyl or heterocyclic groups thatare substituted by the independent replacement of one, two or three ofthe hydrogen atoms thereon with any of the previously mentionedsubstituents. Additionally, it will be appreciated that any of thealicyclic or heterocyclic moieties described above and herein maycomprise an aryl or heteroaryl moiety fused thereto.

The terms “halo” and “halogen”, as used herein, refer to an atomselected from fluorine, chlorine, bromine and iodine.

The term “haloalkyl” denotes an alkyl group, as defined above, havingone, two, or three halogen atoms attached thereto and is exemplified bysuch groups as chloromethyl, bromoethyl, trifluoromethyl, and the like.

The term “amino”, as used herein, refers to a primary (—NH₂), secondary(—NHR_(x)), tertiary (—NR_(x)R_(y)) or quaternary (—N⁺R_(x)R_(y)R_(z))amine, where R_(x), R_(y) and R_(z) are independently an aliphatic,alicyclic, heteroaliphatic, heterocyclic, aromatic or heteroaromaticmoiety, as defined herein. Examples of amino groups include, but are notlimited to, methylamino, dimethylamino, ethylamino, diethylamino,diethylaminocarbonyl, methylethylamino, iso-propylamino, piperidino,trimethylamino, and propylamino.

The term “acyl”, as used herein, refers to a group having the generalformula —C(═O)R_(b), where R_(b) is an aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic or heteroaromatic moiety, asdefined herein.

As used herein, the terms “aliphatic”, “heteroaliphatic”, “alkyl”,“alkenyl”, “alkynyl”, “heteroalkyl”, “heteroalkenyl”, “heteroalkynyl”,and the like encompass substituted and unsubstituted, saturated andunsaturated, and linear and branched groups. Similarly, the terms“alicyclic”, “heterocyclic”, “heterocycloalkyl”, “heterocycle” and thelike encompass substituted and unsubstituted, and saturated andunsaturated groups. Additionally, the terms “cycloalkyl”,“cycloalkenyl”, “cycloalkynyl”, “heterocycloalkyl”,“heterocycloalkenyl”, “heterocycloalkynyl”, “aromatic”,“heteroaromatic”, “aryl”, “heteroaryl” and the like encompass bothsubstituted and unsubstituted groups.

By the term “protecting group”, as used herein, it is meant that aparticular functional moiety, e.g., O, S, or N, is temporarily blockedso that a reaction can be carried out selectively at another reactivesite in a multifunctional compound. In preferred embodiments, aprotecting group reacts selectively in good yield to give a protectedsubstrate that is stable to the projected reactions; the protectinggroup must be selectively removed in good yield by readily available,preferably nontoxic reagents that do not attack the other functionalgroups; the protecting group forms an easily separable derivative (morepreferably without the generation of new stereogenic centers); and theprotecting group has a minimum of additional functionality to avoidfurther sites of reaction. Oxygen, sulfur, nitrogen and carbonprotecting groups may be utilized. For example, oxygen protecting groupsinclude, but are not limited to, methyl ethers, substituted methylethers (e.g., MOM (methoxymethyl ether), MTM (methylthiomethyl ether),BOM (benzyloxymethyl ether), PMBM or MPM (p-methoxybenzyloxymethylether), to name a few), substituted ethyl ethers, substituted benzylethers, silyl ethers (e.g., TMS (trimethylsilyl ether), TES(triethylsilylether), TIPS (triisopropylsilyl ether), TBDMS(t-butyldimethylsilyl ether), tribenzyl silyl ether, TBDPS(t-butyldiphenyl silyl ether), to name a few), esters (e.g., formate,acetate, benzoate (Bz), trifluoroacetate, dichloroacetate, to name afew), carbonates, cyclic acetals and ketals. In certain other exemplaryembodiments, nitrogen protecting groups are utilized. Nitrogenprotecting groups include, but are not limited to, carbamates (includingmethyl, ethyl and substituted ethyl carbamates (e.g., Troc), to name afew) amides, cyclic imide derivatives, N-Alkyl and N-Aryl amines, iminederivatives, and enamine derivatives, to name a few. It will beappreciated that the present invention is not intended to be limited tothese protecting groups; rather, a variety of additional equivalentprotecting groups can be readily identified using the above criteria andutilized in the present invention. Additionally, a variety of protectinggroups are described in “Protective Groups in Organic Synthesis” T. W.Greene and P. G. Wuts (Eds.), John Wiley & Sons: New York, 1999 (3^(rd)Ed), the entire contents of which are incorporated herein by reference.

As used herein, the term “medicament” refers to any substance orcombination of substances that has a beneficial or therapeutic effect.In preferred embodiments of the present invention, the manufacture of amedicament comprises the use of at least one derivative of therocaglamide/aglain/forbaglin family prepared by the methods providedherein. For example, a medicament according to the present invention maycomprise one or more derivatives of the rocaglamide/aglain/forbaglinfamily as active ingredient(s). A medicament may further comprise one ormore other active ingredients, such as drugs or therapeutic agents knownin the art or newly discovered agents whose activity is to be tested,and/or one or more pharmaceutically acceptable carriers. As used herein,the term “pharmaceutically acceptable carrier” refers to a carriermedium which does not interfere with the effectiveness of the biologicalactivity of the active ingredients and which is not excessively toxic tothe hosts at the concentrations at which it is administered. The termincludes solvents, dispersion media, coatings, antibacterial andantifungal agents, isotonic agents, absorption delaying agents, and thelike. The use of such media and agents for pharmaceutically activesubstances is well known in the art (see, for example, Remington'sPharmaceutical Sciences, E. W. Martin, 18^(th) Ed., 1990, MackPublishing Co.: Easton, Pa.).

The term “treatment” is used herein to characterize a method or processthat is aimed at (1) delaying or preventing the onset of a disease orcondition; or (2) slowing down or stopping the progression, aggravation,or deterioration of the symptoms of the disease or condition; or (3)bringing about ameliorations of the symptoms of the disease orcondition; or (4) curing the disease or condition. The treatment may beadministered prior to the onset of the disease, for a prophylactic orpreventive action. Alternatively or additionally, the treatment may beadministered after initiation of the disease or condition, for atherapeutic action.

DETAILED DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS

The present invention is directed to a new, unified biomimetic approachto the synthesis of rocaglamides and related aglains and forbaglins. Anembodiment of this new approach is outlined in FIG. 3.

More specifically, the inventive synthetic method shown in FIG. 3involves photochemical generation of an oxidopyrylium species (compound7) via excited state intramolecular proton transfer (ESIPT) of a3-hydroxyflavone derivative 6 followed by 1,3-dipolar cycloaddition(i.e., [3+2] cycloaddition) of the oxidopyrylium species to adipolarophile, such as a cinnamate derivative. This reaction results inthe formation of the adduct 8, which contains an aglain core structure.Conversion of 8 by oxidative cleavage yields forbaglin 9, whilereduction of the adduct 8 produces aglain 10. Core structure 8 mayalternatively be converted to hydrorocaglate 11 by α-ketol (acyloin)rearrangement; and hydroxyl-directed reduction of 11 affords rocaglate12.

I. Excited State Intramolecular Proton Transfer (ESIPT)

An ESIPT phenomenon involves a very fast intramolecular transfer of aproton. In some cases, this process takes place in only tens or hundredsof femtoseconds (M. Kasha, J. Chem. Soc. Faraday Trans. 2, 1986, 82:2379-2392; B. J. Schwartz et al., J. Phys. Chem., 1992, 96: 3591-3598;F. Laermer et al., Chem. Phys. Lett., 1988, 148: 119-124).

Literature reports have documented excited state intramolecular protontransfer (see, for example, P.-T. Chou, J. Chin. Chem. Soc., 2001, 48:651-682; A. D. Roschal et al., J. Phys. Chem. A, 1998, 102: 5907-5914;A. Bader et al., J. Phys. Chem. A, 2002, 106: 2844-2849 and referencestherein; A. Samanta et al., J. Phys. Chem. A, 2003; 107: 6334-6339; A.P. Demchenko, J. Phys. Chem. A, 2003, 107: 4211-4216; R. Rastogi et al.,Spectrochem. Acta, Part A, 2001, 57: 299-308) of 3-hydroxyflavonederivatives leading to the formation of an oxidopyrylium species (J.Hendrickson and J. S. Farina, J. Org. Chem., 1980, 45: 3359-3361; P. G.Sammes et al., J. Chem. Soc. Perkin Trans. I, 1983, 1261-1265; P. A.Wender et al., J. Am. Chem. Soc., 1997, 119: 12976-12977; J. E. Baldwinet al., Tetrahedron Lett., 2003; 44: 4543-4545).

The overall ESIPT process (shown on FIG. 4 in the case of the parentmolecule, 3-hydroxyflavone, 3-HF) involves generation of a putativetautomeric form of 3-HF, where the proton of the hydroxyl group atposition C3 migrates to the ketone group at position C4 to give anoxidopyrylium species (tautomeric form T).

Although ESIPT processes of 3-HF derivatives have been reported in theliterature to produce excited state species such as the oxidopyrylium,there are no reports of chemical reactions using these species. Thepresent invention encompasses the recognition by the Applicants that thereactivity of such oxidopyrylium species can be advantageously exploitedin chemical reactions.

Accordingly, one aspect of the present invention relates to the use ofphotochemically generated oxidopyrylium species as intermediates inchemical reactions. Preferably, the oxidopyrylium species isphotochemically generated via a process comprising an excited stateintramolecular proton transfer.

As will be appreciated by those of ordinary skill in the art, anyorganic molecule which can produce an oxidopyrylium species uponphotochemical excitation is suitable for use in the practice of thepresent invention. Particularly suitable compounds comprise a5-hydroxy-pyran-4-one group/motif, including, but not limited to,5-hydroxy-2,3-dihydro-pyran-4-one derivatives. 3-hydroxychromonederivatives (M. Itoh, Pure and Applied Chemistry, 1993, 65: 1629-1634;A. S. Klymchenko et al., New J. Chem., 2004, 28: 687-692) and3-hydroxyflavone derivatives. When the photochemically generatedoxidopyrylium species is used in the preparation of rocaglamides andrelated aglains and forbaglins according to the new synthetic approachprovided herein, the oxidopyrylium species is preferably generated byphotochemical excitation of a 3-hydroxychromone derivative of chemicalstructure (I) or 3-hydroxyflavone derivative of chemical structure (II).

Methods for photochemically exciting organic molecules are known in theart. Photochemical irradiation of 3-hydroxyflavone derivatives isdescribed in Example 1 and Example 5.

II. Cycloaddition Reactivity of Oxidopyrylium Species

In preferred embodiments, the photochemically generated oxidopyryliumspecies is used as a reactive intermediate in a cycloaddition, such as a1,3-dipolar cycloaddition.

Initial efforts by the Applicants toward understanding the cycloadditionreactivity of the oxidopyrylium species T (see FIG. 4) were focused onmodel studies using 3-hydroxyflavone, the parent compound and simplestmolecule of the 3-hydroxyflavone family.

Oxidopyrylium Species Generated from 3-Hydroxyflavone

Photoirradiation of 3-hydroxyflavone 13 in presence of the dipolarophilemethyl cinnamate 14 was carried out in acetonitrile using a 450 Wpressure mercury lamp (uranium filter, λ>350 nm). After irradiation atroom temperature for 2 hours, compound 13 was consumed and a mixture ofproducts was obtained, resulting, presumably, from [3+2] cycloaddition(see FIG. 5 and Example 1).

Based on spectroscopic data and X-ray analysis of a crystallinederivative (see Example 1), the major compound (56%) was confirmed to bethe endo cycloadduct 16 in which the phenyl ring of the dipolarophile isanti to the oxido bridge (P. G. Sammes and L. J. Street, J. Phys. Chem.,1998, 102: 5907-5914). ¹H-NMR and ¹³C-NMR spectra recorded for compound16 are presented in FIG. 6. Interestingly, an equilibrium between 16 andthe benzo[b]cyclobutapyran-8-one 17 is observed during silica gelpurification resulting from an acid-mediated ketol shift (X. Creary etal., J. Org. Chem., 1985, 50: 1932-1938). The equilibrium between thetwo core structures was found to be controlled by temperature: heating amixture of compounds 16 and 17 (ethyl acetate, 65° C.) was observed tolead to the formation of compound 16 exclusively. Monitoring of thephotocycloaddition by ¹H-NMR (in CD₃CN) also confirmed formation of 16as the major product (see FIG. 7 and FIGS. 8(A and B)).

Compound 15 (14%) was identified as a cyclopenta[b]tetrahydrobenzofuranby further conversion into a crystalline derivative. In contrast to 16,compound 15 is derived from exo [3+2] cycloaddition to an aglaforbesintype ring system (see compound 4 in FIG. 1) followed by acycloinrearrangement during the photoirradiation process (further experimentsto support the ESIPT mechanism were conducted using 3-methoxyflavone).Irradiation (350 nm, acetonitrile, 5 equivalents of 18, at roomtemperature) did not give a [3+2] cycloadduct but instead provided aproduct resulting from oxidative photocycloaddition (T. Matsuura and T.Takemo, Tetrahedron, 1973, 3337-3340).

Conversion of Cycloadduct 16

Cycloadduct 16, which contains an aglain core structure, was thenevaluated for its ability to be converted to compounds containingrocaglamide and forbaglin ring systems (as shown on FIG. 9). Oxidativecleavage of the aglain core to the forbaglin ring system may beconducted using Pb(OAc)₄ (E. Baer, J. Am. Chem. Soc., 1940, 62:1597-1606). Treatment of cycloadduct 16 with Pb(OAc)₄ inbenzene/methanol at room temperature afforded benzo[b]oxepines 18:19 asa 2:1 mixture of keto-enol tautomers (85%) (see Example 2).

The aglain core structure of compound 16 may alternatively be convertedto dehydrorocaglate by α-ketol (acyloin) rearrangement (L. A. Paquetteand J. E. Hofferberth, Org. React., 2003, 62: 477-567; for ketol shiftsin biogenesis, see, for example, M. Rentzea and E. Hecker, TetrahedronLett., 1982, 23: 1785-1788; and D. H. G. Crout and D. L. Rathbone, J.Chem. Soc. Chem. Commun., 1987, 290-291)

Attempted thermal acycloin rearrangement (J. Lui et al., Tetrahedron,1998, 54: 11637-11650) of compound 16 did not afford any observableketol shift product. Acyloin rearrangements have alternatively beenconducted using acidic or basic conditions or employing metal catalysisand have been used with success in a number of natural product syntheses(for K252a, see, for example, K. Tamaki et al., Tetrahedron Lett., 2002,43: 379-382; for Taxanes, see, for example, L. Paquette and J. E.Hofferberth, J. Org. Chem., 2003, 68: 2266-2275).

Treatment of cycloadduct 16 with protic or Lewis acidic conditions (BF3,Et₂O, ZnCl₂) resulted in decomposition of the starting material.However, treatment of cycloadduct 16 under basic conditions (2.5equivalents of NaOMe, methanol) (X. Creary et al., J. Org. Chem., 1985,50: 1932-1938), afforded a 1:1 mixture of keto-enol tautomers 20:21 (seeExample 3). The success of basic conditions for α-ketol rearrangementmay be explained by the fact that such basic conditions favor theformation of the enolate of 21, which may drive the ketol shiftequilibrium (E. Piers et al., Synlett., 1999, 7: 1082-1084) towards therocaglamide core.

Further proof for this assumption was provided by treatment ofcycloadduct 16 with NaH (2.1 equivalent, tetrahydrofuran, roomtemperature) and quenching of the reaction mixture with thionylchloride, which led to the formation of the stable 1,3,2-dioxathiolane22 (48%) (M. Shipman et al., Tetrahedron, 1999, 55: 108445-10850) (seeExample 3).

Hydroxyl-directed reduction (B. Trost et al., J. Am. Chem. Soc., 1990,112: 9022-9024) of 20:21 afforded rocagolate 23 (95%) (see Example 4).The ¹H-NMR and ¹³C-NMR spectra of compound 23 are presented on FIG. 10.

Oxidopyrylium Species Generated from Methoxy-Substituted3-Hydroxyflavone

3-Hydroxyflavone derivatives with methoxy substitutions were thenevaluated for their suitability in the synthesis of rocaglamides andrelated compounds. The overall synthetic scheme is presented on FIG. 11in the case of the trimethoxy-substituted 3-hydroxyflavone.Trimethoxy-substituted 3-hydroxyflavone was synthesized following aprocedure adapted from a reaction sequence reported by H. Tanaka andcoworkers (Tetrahedron Lett., 2000, 41: 9735-9739) as shown in FIG. 12.Photoirradiation (uranium filter) of kaempferol derivative 24 and methylcinnamate 14 (Y.-J. Lee and T.-D. Wu, J. Chin. Chem. Soc., 2001, 48:201-206) in methanol at 0° C. afforded the aglain 25 as well asbenzo[b]cyclobutapyran-8-one 26 (33% and 17%, respectively) afterpurification on SiO₂ (see Example 5).

Conversion of Compounds 25 and 26

Basic conditions (NaOMe, methanol) were used to effect α-ketolrearrangement of compound 25 and compound 26 (see Example 6). In thecase of compound 25, the reaction led to the formation of a mixture ofendo and exo cycloadducts 27, in which the endo isomer was obtained as amixture of keto-enol tautomers 27′/27″ (the chemical structures ofcompounds 27, 27′ and 27″ are presented on FIG. 12). In the case ofcompound 26, the base-mediated reaction only gave the endo cycloadduct27.

Hydroxyl-directed reduction of keto rocaglate 27, which is described inExample 7, afforded (±)-methyl rocaglate 28 (51%) and the correspondingexo stereoisomer 29 (27%) (B. Trost et al., J. Am. Chem. Soc., 1990,112: 9022-9024). The ¹H-NMR and ¹³C-NMR spectra of compounds 28 and 29are reported in FIG. 14 and FIG. 15, respectively. Spectral data forsynthetic compound 28 were in full agreement with those reported fornatural methyl rocaglate (F. Ishibashi et al., Phytochemistry, 1993, 32:307-310) (see Example 7). Similarly, spectral data for synthetic 29 werein full agreement with those reported for natural methyl rocaglate (G.A. Kraus and J. O. Sy, J. Org. Chem., 1989, 54: 77-83).

Methyl cinnamate was used as dipolarophile in most of the experimentsreported herein. However, as will be appreciated by those skilled in theart, any dipolarophile exhibiting reactivity toward a photochemicallygenerated oxidopyrylium species can be used in the practice of thesynthetic methods disclosed herein.

III. Chemical Modifications of Aglain/Rocaglamide/Forbaglin Derivatives

As will be appreciated by those of ordinary skill in the art, initiallyformed aglain derivatives as well as the forbaglins and rocaglamidesderived from them can be further chemically modified to obtain newderivatives of the aglain/rocaglamide/forbaglin family.

For example, chemical modifications may be performed to studystructure-activity relationships with the goal of developing compoundsthat possess improved biological activity and that fulfill allstereoelectronic, physicochemical, pharmacokinetic, and toxicologicfactors required for clinical usefulness. In such studies, molecularstructure and biological activity are correlated by observing theresults of systemic structural modification on defined biologicalendpoints. For example, comparison of the activity ofstructurally-related compounds may help identify positions and/orchemical motifs that play an important role in biological activity.Similarly, analysis of the effects of the stereochemistry (i.e., thearrangement of atoms in space) of these chemically modified compounds onbiological endpoints may help identify conformations that are favorableto the biological activity. The present invention is intended toencompass chemically modified derivatives of theaglain/rocaglamide/forbaglin family obtained by the methods disclosedherein.

Examples of such chemical modifications are described in Examples 8 and9 in the case of compounds 16 and 15, respectively. The chemicalstructures of the products of these chemical modifications (compound 30and compound 31, respectively) are shown on FIG. 19.

IV. Uses of Aglain/Rocaglamide/Forbaglin Derivatives

As mentioned above, compounds in the rocaglamide/aglain/forbaglin familyhave been demonstrated to exhibit biological activity. In particular, anumber of these compounds are potent natural insecticides (B. W. Nugrohoet al., Phytochemistry, 1997, 45: 1579-1585; B. W. Gussregen et al.,Phytochemistry, 1997, 44: 1455-1461; G. Brader et al., J. Nat. Prod.,1998, 61: 1482-1490; J. Hiort Chaidir et al., Phytochemistry, 1999, 52:837-842; B. W. Nugrobo et al., Phytochemistry, 1999, 51: 367-376).

Moreover, rocaglamide derivatives have been found to exhibit cytostaticactivity in human cancer cell lines (B. Cui et al., Tetrahedron, 1997,53: 17625-17632; T. S. Wu et al., J. Nat. Prod., 1997, 60: 606-608; S.K, Lee et al., Chem. Biol. Interact., 1998, 115: 215-228) with effectscomparable to those observed with established anticancer drugs such asvinblastine sulfate and actinomycin D (F. I. Bohnenstengel et al., Z.Naturforsch. [C], 1999, 54: 55-60; F. I. Bohnenstengel et al., Z.Naturforsch. [C], 1999, 54: 1075-1083). In particular, the rocaglatesilvestrol 2 (see FIG. 1) has been shown to display cytotoxic activityagainst human cancer cells comparable to the anticancer drug Taxol (B.Y. Hwang et al., J. Org. Chem., 2004, 69: 3350-3358). Other studies haverevealed that these compounds block cell cycle progression and induceapoptosis at nanomolar concentrations in colorectal tumor cell lines (B.Hausott et al., Int. J. Cancer, 2004, 109: 933-940). Experimentalresults reported in this study suggest that apoptosis is induced via ap38-mediated stress pathway (B. Hausott et al., Int. J. Cancer, 2004,109: 933-940). Furthermore, rocaglamides have been demonstrated to blockprotein biosynthesis (T. Ohse et al., J. Nat. Prod., 1996, 650-653) andto induce growth arrest in the G2/M phase in certain tumor cells (F. I.Bohnenstengel et al., Z. Naturforsch. [C], 1999, 54: 1075-1083).

More recently, it was shown that rocaglamides represent highly potentand specific inhibitors of TNF-α (tumor necrosis factor-alpha) and PMA(porbol 12-myristate 13 acetate)-induced NF-κB (nuclear factor-kappa B)activity in different mouse and human T cell lines. The IC₅₀ valuesobserved for rocaglamide derivatives were in the nanomolar range whereasaglain derivatives proved inactive. Rocaglamide and several of itsderivatives are among the strongest inhibitors of NF-κB-induced geneactivation known so far (B. Baumann et al., J. Biol. Chem., 2002, 277:44791-44800).

Agents that can suppress NF-κB activation have, in principle, thepotential to prevent or delay onset or treat NF-κB-linked diseases. Onactivation, NF-κB induces the expression of more than 200 genes, thathave been shown to suppress apoptosis, induce cellular transformation,proliferation, invasion, metastasis, chemoresistance, radioresistance,and inflammation (A. Garg and B. B. Aggarwal, Leukemia, 2002, 16:1053-1056). The activated form of NF-κB has been found to mediate cancer(A. Garg and B. B. Aggarwal, Leukemia, 2002, 16: 1053-1056; A Lin and M.Karin, Semin. Cancer Biol., 2003, 13: 107-114; R. Z. Orlowski and A. S.Baldwin, Trends Mol. Med., 2002, 8: 385-389), atherosclerosis (G. Valenet al., J. Am. Coll. Cardiol., 2001, 38: 307-314), myocardial infarction(W. K. Jones et al., Cardiovasc. Toxicol., 2003, 3: 229-254), diabetes(S. E. Shoelson et al., Int. J. Obes. Relat. Metab. Disord., 2003,27(Suppl. 3): S49-52), allergy (L. Yang et al., J. Exp. Med., 1998, 188:1739-1750; J. Das et al., Nature Immunol., 2001, 2: 45-50), asthma (R.Gagliardo et al., Am. J. Respir. Crit. Care Med., 2003, 168: 1190-1198),arthritis (A. K. Roshak et al., Curr. Opin. Pharmacol., 2002, 2:316-321), Crohn's disease (D. A. van Heel et al., Hum. Mol. Genet.,2002, 11: 1281-1289), multiple sclerosis (C. J. Huang et al., Int. J.Dev. Neurosci., 2002, 20: 289-296), Alzheimer's disease (M. P. Mattsonand S. Camandola, J. Clin. Invest., 2001, 107: 247-254; B. Kaltschmidtet al., Proc. Natl. Acad. Sci. USA, 1997, 94: 2642-2647), osteoporosis,psoriasis, septic shock, AIDS and other inflammatory diseases (J. R.Burke, Curr. Opin. Drug Discov. Devel., 2003, 6: 720-728; Y. Yamamotoand R. B. Gaynor, Curr. Mol., Med., 2001, 1: 287-296; Y. Yamamoto and R.B. Gaynor, J. Clin. Invest., 2001, 107: 135-142).

Interestingly, a synthetic derivative of the natural product rocaglaolwas recently found to exhibit neuroprotective activity in vitro and inanimal models of Parkinson's disease and traumatic brain injury (T.Fahrig et al., Mol. Pharmacol., (Fast Forward” publications), Feb. 16,2005). Experimental data reported in this study suggest that byinhibiting NF-κB and AP-1 (activator protein-1) signaling, thisrocaglaol derivative is able to reduce tissue inflammation and neuronalcell death resulting in significant neuroprotection in animal models ofacute and chronic neurodegeneration.

Accordingly, another aspect of the present invention relates to the useof derivatives of the rocaglamide/aglain/forbaglin family for themanufacture of medicaments for use in the treatment of various diseasestates, including cancer and cancerous conditions, conditions associatedwith cellular hyperproliferation, and NF-κB-associated conditions.Preferably, the rocaglamide derivatives used in the manufacture of thesemedicaments are prepared by the inventive methods disclosed herein.

Cancer and cancerous conditions that can be treated using suchmedicaments may be selected from the group consisting of leukemia,sarcoma, breast, colon, bladder, pancreatic, endometrial, head and neck,mesothelioma, myeloma, oesophagal/oral, testicular, thyroid, cervical,bone, renal, uterine, prostate, brain, lung, ovarian, skin, liver andbowel and stomach cancers, tumors and melanomas. Conditions associatedwith cellular hyperproliferation that can be treated using the inventivemedicaments may be selected from the group consisting ofatherosclerosis, restenosis, rheumatoid arthritis, osteoarthritis,inflammatory arthritis, psoriasis, periodontal disease and virallyinduced cellular hyperproliferation. NF-κB associated conditions thatcan be treated using the medicaments disclosed herein may be selectedfrom the group consisting of immunological disorders, septic shock,transplant rejection, radiation damage, reperfusion injuries afterischemia, arteriosclerosis and neurodegenerative diseases.

The medicaments according to the present invention may be in a liquid,aerosol or solid dosage form, and may be manufactured into any suitableformulation including, but not limited to, solutions, suspensions,micelles, emulsions, microemulsions, syrups, elixirs aerosols,ointments, gels suppositories, capsules, tablets, pills, dragees, andthe like, as will be required for the appropriate route ofadministration.

Any suitable route of administration of the inventive medicaments isencompassed by the present invention including, but not limited to,oral, intravenous, intraperitoneal, intramuscular, subcutaneous,inhalation, intranasal, topical, rectal or other administration routeknown in the art. The route of administration, formulation and dosage ofthe medicament will be dependent upon a variety of factors including thepathophysiological condition to be treated and the severity and/orextent of the disorder, the age, sex, weight and general health of theparticular patient, the potency, bioavailability, in vivo half-life andseverity of the side effects of the specific rocaglamide derivative(s)employed in the manufacture of the medicament, the time ofadministration, the duration of the treatment, drugs used in combinationor coincidental with the specific rocaglamide derivative(s) employed,and similar factors well known in the art. These factors are readilydetermined in the course of therapy. Alternatively or additionally, thedosage to be administered can be determined from studies using animalmodels for the particular condition to be treated, and/or from animal orhuman data obtained for compounds which are known to exhibit similarpharmacological activities. A medicament may be formulated in such a waythat the total dose required for each treatment is administered bymultiple dose or in a single dose. In certain embodiments, themedicament is manufactured or formulated in dosage unit form. Theexpression “dosage unit form”, as used herein, refers to a physicallydiscrete unit of medicament appropriate for the condition/patient to betreated.

In certain embodiments, a medicament according to the present inventioncomprises one or more rocaglamide derivatives as active ingredients. Inother embodiments, the medicament further comprises one or more othertherapeutic agents. The nature of such additional therapeutic agent(s)will depend on the condition to be treated by administration of themedicament. The ability to determine combinations of compounds suitableto treat particular disorders is well within the capabilities of trainedscientists or physicians. For example, a medicament according to thepresent invention for use in the treatment of cancer may furthercomprise approved chemotherapeutic drugs, including, but not limited to,alkylating drugs (mechlorethamine, chlorambucil, Cyclophosphamide,Melphalan, Ifosfamide), antimetabolites (Methotrexate), purineantagonists and pyrimidine antagonists (6-Mercaptopurine,5-Fluorouracil, Cytarabile, Gemcitabine), spindle poisons (Vinblastine,Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (Etoposide,Irinotecan, Topotecan), antibiotics (Doxorubicin, Bleomycin, Mitomycin),nitrosoureas (Carmustine, Lomustine), inorganic ions (Cisplatin,Carboplatin), enzymes (Asparaginase), and hormones (Tamoxifen,Leuprolide, Flutamide, and Megestrol), to name a few. For a morecomprehensive discussion of updated cancer therapies see,http://www.nci.nih.gov/, a list of the FDA approved oncology drugs athttp://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck Manual,7th Ed. 1999, the entire contents of which are hereby incorporated byreference.

In addition to the active ingredient(s), the medicament may furthercomprise one or more pharmaceutically acceptable carriers, including,but not limited to, inert diluents, dispersion media, solvents,solubilizing agents, suspending agents, emulsifying agents, wettingagents, coatings, isotonic agents, sweetening, flavoring and perfumingagents, antibacterial and antifungal agents, absorption delaying agents,and the like. The use of such media and agents for the manufacture ofmedicaments is well known in the art (see, for example, Remington'sPharmaceutical Sciences, E. W. Martin, 18^(th) Ed., 1990, MackPublishing Co., Easton, Pa.).

EXAMPLES

The following examples describe some of the preferred modes of makingand practicing the present invention. However, it should be understoodthat these examples are for illustrative purposes only and are not meantto limit the scope of the invention. Furthermore, unless the descriptionin an Example is presented in the past tense, the text, like the rest ofthe specification, is not intended to suggest that experiments wereactually performed or data were actually obtained.

The novel biomimetic approach to the synthesis of rocaglamides, aglainsand forbaglins has recently been described, by the Applicants, in ascientific article (B. Gerard et al., J. Am. Chem. Soc., 2004, 126:13620-13621), which is incorporated herein by reference in its entirety.

General Information

Melting points were recorded on a MeI-Temp (Laboratory Devices). Yieldsrefer to chromatographically and spectroscopically pure materials,unless otherwise stated. Methylene chloride, acetonitrile, methanol, andbenzene were purified by passing through two packed columns of neutralalumina (Glass Contour, Irvine, Calif.). 3-Hydroxyflavone was purchasedfrom Indofine Chemical Company, Inc. (Hillsborough, N.J.).

Nuclear Magnetic Resonance. ¹H-NMR spectra were recorded at 400 MHz atambient temperature with CDCl₃ as solvent unless otherwise stated.¹³C-NMR spectra were recorded at 75.0 MHz at ambient temperature withCDCl₃ as solvent unless otherwise stated. Chemical shifts are reportedin parts per million (ppm) relative to CDCl₃ (¹H, δ 7.24; ¹³C, δ 77.0)or acetone-d₆ (1H, δ 2.04; ¹³C, δ 207.6, 30.0). Data for ¹H-NMR arereported as follows: chemical shift, integration, multiplicity(abbreviations are as follows: app=apparent, par obsc=partially obscure,ovrlp=overlapping, s=singlet, d=doublet, t=triplet, q=quartet,m=multiplet) and coupling constants. All ¹³C-NMR spectra were recordedwith complete proton decoupling.

Infrared Spectroscopy. Infrared spectra were recorded on a Nicolet Nexus670 FT-IR spectrophotometer. Low and high-resolution mass spectra wereobtained at the Boston University Mass Spectrometry Laboratory using aFinnegan MAT-90 spectrometer.

Chromatography. HPLC analyses were carried out on an Agilent 1100 seriesHPLC (CHIRALCEL OD, Column No. OD00CE-AI015 and Agilent Zorbax SB-C18).Analytical thin layer chromatography was performed using 0.25 mm silicagel 60-F plates; and flash chromatography, using 200-400 mesh silica gel(Scientific Absorbents, Inc.).

Photochemical Irradiation. Photochemical irradiation experiments wereperformed using a Hanovia 450 W medium pressure mercury lamp housed in awater-cooled quartz immersion well or using an ethylene glycol coolingsystem (Neslab, RTE-140). Pyrex test tubes (16×100 mm) were mounted on asupport approximately 0.5 cm from the immersion well lamp. An uraniumfilter was obtained from James Glass (Hanover, Mass.).

All other reactions were carried out in oven-dried glassware under anargon atmosphere unless otherwise noted.

Example 1 Photochemical Irradiation of 3-Hydroxyflavone Irradiation of3-Hydroxyflavone in the Presence of Methyl Cinnamate

To a (16×100 mm) test tube was added 3-hydroxyflavone 13 (400 mg, 1.7mmol) and methyl cinnamate 14 (650 mg, 4 mmol) in 8 mL of anhydrousacetonitrile. After degassing with argon for 5 minutes, the mixture wasirradiated (Hanovia UV lamp uranium filter, water used for cooling) atroom temperature for 2 hours. The solution was concentrated in vacuo toafford a pink-yellow oil.

Purification via flash chromatography (60:40 hexanes/EtOAc) yielded 92mg (0.23 mmol, 15%) of cyclopenta[b]tetrahydrobenzofuran 15 and 370 mg(0.94 mmol, 56%) of a mixture of cyclopenta[bc]benzopyran 16 andbenzo[b]cyclobutapyran-8-one 17 as colorless solid. Compound 17 wasquantitatively converted to cyclopenta[bc]benzopyran 16 by thermolysis(EtOAc, 65° C., 4 hours).

Cyclopenta[b]tetrahydrobenzofuran 15. White solid: mp 76-78° C.; IRν_(max) (film): 3449, 3064, 3033, 2955, 2920, 1740, 1697, 1682, 1596,1476, 1254, 1223, 755 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.46-6.97 (14H,m), 4.48 (1H, d, J=13 Hz), 3.96 (1H, d, J=13 Hz), 3.59 (3H, s), 3.01(1H, s) ppm; ¹³C-NMR (75 MHz, CDCl₃): 208.9, 168.8, 159.6, 136.9, 134.9,132.1, 129.1, 129.0, 128.9, 128.3, 134.9, 132.1, 129.1, 129.0, 128.9,128.3, 127.9, 126.5, 125.8, 124.8, 122.5, 110.7, 94.0, 87.8, 59.3, 52.4,52.3 ppm; HRMS (EI) m/z calculated for C₂₅H₂₀O₅, 400.1311; found,401.1429 (M+H).

Cyclopenta[bc]benzopyran 16. White solid: mp 78-81° C.; IR ν_(max)(film): 3452, 3060, 3033, 2940, 1767, 1736, 1608, 1584, 1483, 1452,1210, 905 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.34-7.82 (14H, m), 4.631 (1H,d, J=9.2 Hz), 3.645 (1H, d, J=9.2 Hz), 3.606 (3H, s), 3.57 (1H, s) ppm;¹³C-NMR (75 MHz, CDCl₃) δ 208.4, 170.1, 150.9, 138.2, 133.4, 130.8,129.8, 128.9, 128.7, 128.4, 128.0, 127.9, 127.5, 127.4, 127.3, 126.8,126.6, 124.9, 122.1, 116.1, 85.1, 79.8, 57.0, 54.2, 52.8 ppm; HRMS(CI/NH₃) m/z calculated for C₂₅H₂₀O₅, 400.1311; found, 401.1357 (M+H).

Benzo[b]cyclobutapyran-8-one 17. White solid: mp 68-70° C.; IR ν_(max)(film): 3448, 2922, 2851, 1743, 1597, 1558, 1475, 1248, 1055, 998, 965,755 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.63-7.61 (2H, m), 7.25-6.95 (12H,m), 4.25 (1H, d, J=8.8 Hz), 3.74 (1H, d, J=8.8 Hz), 3.55 (3H, s), 3.27(1H, s) ppm; ¹³C-NMR δ 190.33, 169.6, 151.5, 139.4, 135.4, 130.2, 129.9,128.9, 128.7, 128.4, 128.1, 127.8, 127.5, 127.4, 126.8, 124.9, 124.6,121.3, 116.5, 97.5, 88.6, 60.9, 54.3, 52.4 ppm; HRMS (CI/NH₃) m/zcalculated for C₂₅H₂₀O₅, 400.1311; found, 401.1357 (M+H).

Example 2 Conversion of Cycloadduct 16 to a Forbaglin Ring System

50 mg of cyclopenta[bc]benzopyran 16 (0.125 mmol, 1 equiv) weredissolved in a mixture of methanol (30%) and benzene (0.9 mL/2.1 mL).Pb(OAc)₄ (55 mg, 0.125 mmol, 1 equivalent) was then added portionwise atroom temperature and the reaction was stirred for 30 minutes at roomtemperature. After removal of the solvent in vacuo, the resultingresidue was diluted with water (5 mL) and EtOAc (5 mL). After separationof the organic layer, the aqueous layer was further extracted twice withEtOAc (5 mL). The organic extracts were combined, washed with brine,dried over MgSO₄, filtered, and concentrated in vacuo. Purification onsilica gel (20% EtOAc in hexane) afforded 46 mg (0.11 mmol, 85%) of18:19 as a colorless solid (2:1 mixture of keto/enol tautomers.

Benzo[b]oxepines 18/19. Colorless solid: mp 178-181° C.; IR ν_(max)(film): 3060, 3033, 2959, 2924, 1759, 1747, 1684, 1602, 1444, 1434,1308, 1244; 1102 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.64-7.62 (2H, d, J=7.2Hz), 7.44-7.28 (8H, m), 7.18-7.16 (4H, m), 5.12 (1H, d, J=10 Hz), 4.41(1H, d, J=10 Hz), 3.66 (3H, s), 3.16 (3H, s) ppm; ¹³C-NMR (75 MHz,CDCl₃) δ 193.2, 156.7, 154.2, 139.0, 134.8, 132.4, 129.2, 129.1, 128.9,128.7, 128.3, 128.2, 127.8, 127.7, 127.6, 127.4, 126.9, 126.7, 122.3,121.9, 121.8, 121.6, 64.9, 52.5, 52.2, 52.0, 51.8, 49.8, 46.7 ppm; HRMS(CI/NH₃) m/z calculated for C₂₆H₂₂O₆, 430.1416; found, 431.1516 (M+H).

Example 3 Conversion of Cycloadduct 16 to a Dehydrorocaglate Ring System

To a solution of cyclopenta[bc]benzopyran 16 (50 mg, 0.125 mmol, 1equivalent) in MeOH (3 mL) at room temperature was added a solution ofNaOMe (17 mg, 0.31 mmol, 2.5 equivalents) in MeOH (1 mL) at roomtemperature. The resulting solution was stirred for 40 minutes at 65° C.After quenching the reaction with saturated NH₄Cl at room temperature,10 mL of EtOAc was added. The organic layer was separated and washedwith water (2×5 mL) and brine (5 mL), dried over MgSO₄, filtered, andconcentrated in vacuo. Purification on silica gel (20% EtOAc in hexane)afforded 45 mg (0.11 mmol, 90%) of the corresponding rocaglates 20/21 asa white solid.

Cyclopenta[b]tetrahydrobenzofurans 20/21. White solid: mp 141-143° C. IRν_(max) (film): 3066, 3027, 2954, 2923, 2856, 1758, 1730, 1650, 1594,1454, 1279, 1247, 1146, 975 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃, 1:1 mixture ofketo/enol tautomers 20:21) δ 7.52-6.88 (28H, m), 5.28 (1H, s), 4.13 (2H,dd, J=13.6 Hz), 3.63 (3H, s), 3.57 (3H, s), 2.66 (1H, s), 2.10 (1H, s)ppm; ¹³C NMR (75 MHz, CDCl₃) δ 204.3, 167.1, 159.8, 132.6, 131.1, 128.8,128.0, 127.8, 127.7, 127.6, 127.6, 127.3, 127.2, 126.9, 126.8, 126.6,126.2, 125.3, 124.8, 122.6, 121.8, 119.5, 112.4, 110.6, 98.7, 57.4,56.7, 55.8, 52.9, 51.7 ppm; HRMS (EI) m/z calculated for C₂₅H₂₀O₅,400.1311; found, 401.1427 (M+H).

To a solution of NaH (washed with 3×10 mL hexanes, 5 mg, 0.21 mmol, 2.1equivalents) in THF (2 mL) was added a solution ofcyclopenta[bc]benzopyran 16 (40 mg, 0.10 mmol, 1 equivalent) in THF (1mL) at room temperature. The resulting yellow solution was stirred atroom temperature for 30 minutes. After addition of thionyl chloride (15μL, 0.21 mmol, 2.1 equivalents) at room temperature, the mixture wasstirred for another hour and then quenched with saturated aqueousNaHCO₃. 10 mL of EtOAc were then added and the organic layer was washedwith 2×3 mL of water and 3 mL brine. The organic extracts were driedover MgSO₄, filtered, and concentrated in vacuo. Purification on silicagel (5% EtOAc in hexane) afforded 21 mg (0.048 mmol, 48%) of thecorresponding 1,3,2-dioxathiolane 22 as a yellow oil.

1,3,2-Dioxathiolane 22. Yellow oil: IR ν_(max) (film): 3025, 2948, 2913,1716, 1650, 1553, 1243, 1200, 746 cm⁻¹. ¹H-NMR (400 MHz, CDCl₃) δ7.46-7.07 (14H, m), 3.85 (1H, s), 3.72 (3H, s) ppm; ¹³C-NMR δ 190.4,165.4, 144.9, 143.1, 132.9, 132.6, 130.8, 130.3, 130.0, 129.6, 129.2,128.8, 128.7, 128.5, 128.1, 125.6, 124.7, 122.6, 111.1, 52.6, 52.4 ppm;HRMS (EI) m/z calculated for C₂₅H₁₈O₆S, 446.0824; found, 447.0805 (M+H).

Example 4 Conversion of Dehydrorocaglate Ring System to Rocaglate RingSystem

To a solution of 197 mg (0.75 mmol, 6 equivalents) of Me₄NBH(OAc)₃ and68 μL (1.25 mmol, 10 equivalents) of acetic acid in 3 mL of CH₃CN wasadded a solution of 50 mg (0.12 mmol, 1 equivalent) of keto rocaglate 20in 1 mL of CH₃CN. The resulting yellow solution was stirred for 12 hoursat room temperature before being quenched with 2 mL of saturated NH₄Clsolution. The solution was then treated with 1 mL of a 3 M aqueoussolution of sodium/potassium tartrate and stirred at room temperaturefor 30 minutes. The aqueous solution was extracted with CH₂Cl₂ (2×5 mL).The combined organic layers were washed with brine, dried over MgSO₄,filtered, and concentrated in vacuo. Purification on silica gel (20%EtOAc in hexane) afforded 30 mg (0.047 mmol, 95%) of 23 as a whitesolid.

Cyclopenta[b]tetrahydrobenzofuran 23. White solid: mp 176-178° C.; IRν_(max) (film): 3421, 3031, 2925, 1733, 1600, 1476, 1462, 1249, 1102,976 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.41-6.96 (14H, m), 4.84 (1H, d, J=6Hz), 4.50 (1H, d, J=13.6 Hz), 3.99 (1H, dd, J=6, 13.6 Hz), 3.66 (3H, s),2.55 (1H, s), 1.82 (1H, s), ppm; ¹³C-NMR (75 MHz, CDCl₃) δ 171.5, 159.1,136.8, 134.5, 131.4, 127.9, 127.7, 127.6, 127.5, 127.4, 126.8, 126.5,126.3, 121.6, 111.0, 100.8, 93.3, 79.2, 56.0, 52.2, 50.8 ppm; LRMS (ESI+) m/z calculated for C₂₅H₂₂O₅, 402.1467; found, 403.0 (M+H).

Example 5 Photochemical Irradiation of Methoxy-Substituted3-Hydroxyflavone Synthesis of Trimethoxy-Substituted 3-Hydroxyflavone

Trimethoxy-substituted 3-hydroxyflavone 24 was synthesized following aprocedure adapted from a reaction sequence reported by H. Tanaka andcoworkers (Tetrahedron Lett., 2000, 41: 9735-9739). The reaction schemeis presented on FIG. 12.

Irradiation of Trimethoxy-Substituted 3-Hydroxyflavone in the Presenceof Methyl Cinnamate

To a (16×100 mm) test tube was added with kaempferol derivative 24 (200mg, 0.61 mmol), methyl cinnamate 14 (990 mg, 6.1 mmol), and 20 mL ofanhydrous methanol. After degassing with argon, the mixture wasirradiated (Hanovia UV lamp, uranium filter) at 0° C. for 12 hours underan argon atmosphere. The solution was concentrated in vacuo to give ayellow oil. Purification via flash chromatography (60:40 hexanes/EtOAc)afforded 100 mg (0.2 mmol, 33%) of the corresponding trimethoxycyclopenta[bc]benzopyran derivative 25 (mixture of endo/exocycloadducts) as a white solid and 50 mg (0.1 mmol, 17%) ofbenzo[b]cyclo-butapyran-8-one derivative 26 as a yellow solid.

Trimethoxy Cyclopenta[bc]benzopyran 25. White solid: mp 83-85° C. IRν_(max) (film): 3475, 3013, 2943, 2832, 1786, 1737, 1611, 1590, 1510,1450, 1255, 1146, 1094, 828 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.54-7.52(2H, d, J=8.8 Hz), 7.25-7.23 (2H, d, J=8.8 Hz), 7.17-7.49 (2H, m),7.10-7.04 (6H, m), 6.85-6.82 (2H, m), 6.64-6.60 (4H, m), 6.19-6.18 (1H,d, J=2 Hz), 6.18-6.17 (1H, d, J=2 Hz), 6.11-6.10 (1H, d, J=2 Hz),6.08-6.07 (1H, d, J=2 Hz), 4.49-4.47 (1H, d, J=9.2 Hz), 4.191-4.168 (1H,d, J=9.2 Hz), 3.94 (1H, s), 3.84 (3H, s), 3.83 (3H, s), 3.77 (4H, m),3.75 (3H, s), 3.71 (3H, s), 3.66 (4H, m), 3.62 (3H, s), 3.55 (3H, s),3.29 (1H, s) ppm; ¹³C-NMR (70 MHz, CDCl₃) δ 205.5, 170.7, 170.6, 161.9,161.3, 158.8, 158.6, 158.4, 153.6, 152.8, 139.9, 138.1, 130.1, 129.8,128.9, 128.7, 128.2, 127.8, 127.9, 127.0, 126.5, 125.6, 113.6, 112.7,112.6, 107.7, 106.5, 97.9, 95.5, 94.4, 94.3, 93.6, 93.4, 92.7, 88.7,83.6, 81.04, 80.7, 62.4, 57.6, 56.1, 55.9, 55.4, 55.3, 55.1, 54.5, 53.4,52.2, 51.8 ppm; HRMS (CI/NH₃) m/z calculated for C₂₈H₂₆O₈, 490.1628;found, 491.1739 (M+H).

Trimethoxy benzo[b]cyclobutapyran-8-one 26. Yellow solid: mp 79-81° C.IR ν_(max) (film): 3489, 3006, 2948, 2839, 1734, 1729, 1618, 1590, 1516,1461, 1437, 1299, 1200, 1148, 1096, 909 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ7.53 (2H, d, J=8.8 Hz), 7.16 (2H, m), 7.01 (3H, m), 6.64 (2H, d, J=8.8Hz), 6.19 (1H, d, J=2 Hz), 6.08 (1H, d, J=2 Hz), 4.27 (1H, s), 4.17 (1H,d, J=9.6 Hz), 3.84 (4H, m), 3.75 (3H, s), 3.67 (3H, s), 3.56 (3H, s)ppm.

Example 6 Conversion of Aglains 25 and 26 to a Keto Rocaglate RingSystem Conversion of Aglain 25

To a solution of aglain 25 (60 mg, 0.12 mmol, 1 equivalent) in MeOH (4mL) was added a solution of NaOMe (13.2 mg, 0.24 mmol, 2.5 equivalents)in MeOH (1 mL) at room temperature. The resulting solution was stirredfor 40 minutes at 65° C. After quenching the reaction with saturatedNH₄Cl, 10 mL of EtOAc was then added, and the organic layer was washedwith water (2×5 mL) and brine (5 mL). The organic layer was dried overMgSO₄, filtered, and concentrated in vacuo to afford 57 mg (0.12 mmol,95%) of crude ketol shift product 27/27′/27″ as a yellow oil which wasused without further purification (3:1 mixture of endo:exo isomers27′/27″ and 27, see chemical structures of 27, 27′, 27″ on FIG. 13).

Trimethoxy rocaglate 27/27′/27″. Yellow oil: IR ν_(max) (film): 3501,3006, 2947, 2926, 2839, 1762, 1734, 1615, 1513, 1450, 1440, 1255, 1213,1146, 1033, 1076 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃, 1:1 mixture of keto/enoltautomers 27′:27″) δ 7.34-7.32 (2H, d, J=6.8 Hz), 7.20-7.19 (2H, m),7.09-6.86 (15H, m), 6.65 (2H, d, J=8.8 Hz), 6.51 (2H., d, J=6.8 Hz),6.33 (1H, d, J=1.6 Hz), 6.17 (1H, d, J=1.6 Hz), 6.13 (1H, d, J=1.6 Hz),6.12 (1H, d, J=1.6 Hz), 6.05 (1H, d, J=1.6 Hz), 6.00 (1H, d, J=1.6 Hz),4.46 (1H, s), 4.42 (1H, d, J=14.8 Hz), 4.36 (1H, d, J=14.8 Hz), 4.22(1H, d, J=13.6 Hz), 4.04 (1H, d, 13.6 Hz), 3.84 (3H, s), 3.08-3.79 (9H,m), 3.77 (9H, m), 3.70 (6H, m), 3.64 (6H, m), 3.57 (3H, s), 3.30 (1H,s), 3.01 (1H, s) ppm; HRMS (EI) m/z calculated for C₂₈H₂₆O₈, 490.1628;found, 490.9634 (M+H).

Conversion of Aglain 26

Benzo[b]cyclobutapyran-8-one 26 was subjected to the aforementionedconditions using 20 mg (0.041 mmol, 1 equivalent) of 26 in MeOH (2 mL)and NaOMe (5 mg, 0.09 mmol, 2.5 equivalents) in MeOH (1 mL). 18 mg ofcrude ketol shift product 27 (0.036, 90%) was isolated and used withoutfurther purification (only the endo isomer was isolated).

Example 7 Hydroxyl-Directed Reduction of Keto Rocaglate 27Hydroxyl-Directed Reduction of Trimethoxy Keto Rocaglate 27

To a solution of 184 mg (0.70 mmol, 6 equivalents) of Me₄NBH(OAc)₃ and63 μL (1.16 mmol, 10 equivalents) of acetic acid in 3 mL of CH₃CN wasadded a solution of 57 mg (0.12 mmol, 1 equivalent) of 27 in 1 mL ofCH₃CN. The resulting yellow solution was stirred for 12 hours at roomtemperature before being quenched with 2 mL of saturated NH₄Cl. Thesolution was then treated with 1 mL of a 3 M aqueous solution ofsodium/potassium tartrate and stirred at room temperature for 30minutes. The aqueous solution was extracted with CH₂Cl₂ (2×5 mL). Thecombined organic layers were washed with brine, dried over MgSO₄,filtered and concentrated in vacuo. Purification on silica gel (40%EtOAc in hexane) afforded 30 mg (0.030 mmol, 51%) of the correspondingendo methyl rocaglate 28 and 18 mg (0.017 mmol, 27%) of thecorresponding exo methyl rocaglate 29.

Endo Methyl Rocaglate 28. White solid: mp 92-93° C.; R ν_(max) (film):3013, 2954, 2926, 2853, 1734, 1615, 1517, 1457, 1433, 1262, 1195, 1150,1031, 832 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.09 (2H, d, J=9.2 Hz),7.05-7.03 (3H, m), 6.84 (2H, m), 6.65 (2H, d, J=9.2 Hz), 6.27 (1H, d,J=2 Hz), 6.1 (1H, d, J=2 Hz), 5.01 (1H, dd, J=6.4, 1.2 Hz), 4.28 (1H, d,J=14.4 Hz), 3.80 (1H, dd, J=14.4, 6.4 Hz), 3.86 (3H, s), 3.82 (3H, s),3.69 (3H, s), 3.63 (3H, s), 3.50 (1H, s), 1.81 (1H, br) ppm; ¹³C-NMR (75MHz, CDCl₃) δ 170.5, 164.1, 160.9, 158.8, 157.0, 137.0, 129.0, 128.4,127.8, 127.7, 126.5, 112.7, 107.7, 101.9, 93.7, 92.7, 89.5, 79.6, 60.4,55.8, 55.1, 55.0, 51.9, 50.6 ppm; 6 HRMS (CI/NH₃) m/z calculated forC₂₈H₂₈O₈, 492.1784; found, 493.1891 (M+H).

Exo Methyl Rocaglate 29. Foamy yellow: solid mp 84-85° C. IR ν_(max)(film): 3031, 3006, 2958, 2936, 2846, 1730, 1636, 1430, 1307, 1258,1132, 103 cm⁻¹; ¹H-NMR (400 MHz, CDCl₃) δ 7.34 (2H, d, J=8.8 Hz),7.17-1.15 (3H, m), 6.95-6.94 (2H, m), 6.87 (2H, d, J=8.8 Hz), 6.12 (1H,d, J=1.6 Hz), 6.06 (1H, d, J=1.6 Hz), 4.76 (1H, dd, J=10.2, 1.6 Hz),4.02 (1H, d, J=12.8 Hz), 3.82 (3H, s), 3.78 (3H, s), 3.77 (3H, s), 3.60(3H, s), 3.23 (1H, dd, J=12.8, 10.2 Hz), 1.81 (1H, s) ppm; ¹³C-NMR (75MHz, CDCl₃) δ 173.1, 164.1, 162.0, 159.4, 157.9, 135.0, 129.1, 128.4,128.0, 127.3, 119.7, 113.6, 105.1, 99.5, 92.6, 91.4, 88.8, 83.9, 55.8,55.8, 55.4, 54.8, 52.3, 50.9 ppm; HRMS (CI/NH₃) m/z calculated forC₂₈H₂₈O₈, 492.1784; found, 493.1891 (M+H).

The crude ketol shift product 27 obtained frombenzo[b]cyclobutapyran-8-one derivative 26 was subjected to theaforementioned conditions using 58 mg of Me₄NBH(OAc)₃ (0.22 mmol, 6equivalents), 20 μL (0.37 mmol, 10 equivalents) in 3 mL of MeCN, and 18mg (0.037 mmol, 1 equivalent) of compound 26. 13 mg of endo methylrocaglate 28 (0.021 mmol, 75%) was obtained.

Tables 1, 2, and 3 shown below summarize data comparison of natural (F.Ishibashi et al., Phytochemistry, 1993, 32: 307-310) and synthetic endomethyl rocaglate 28.

TABLE 1 ¹H-NMR Data (400 MHz, CDCl₃) for natural and synthetic endomethyl rocaglate 28. ¹H-NMR (400 Hz in CDCl₃) Position Natural Synthetic28 1 5.02 (dd, 1.6, 6.8) 5.01 (dd, 1.2, 6.4) 2β 3.91 (dd, 6.8, 14.4)3.91 (dd, 6.4, 14.4) 3α 4.32 (d, 14.4) 4.27 (d, 14.4) 5 6.29 (d, 2.4)6.26 (d, 2) 7 6.13 (d, 2.4) 6.10 (d, 2) 2′, 6′ 7.11 (d, 8.8) 7.10 (d,9.2) 3′, 5′ 6.68 (d, 8.8) 6.65 (d, 9.2) 2″, 6″ 6.88 (m) 6.85 (m) 3″, 4″,5″ 7.07 (m) 7.04 (m) OMe-6 3.88 (s) 3.86 (s) OMe-8 3.84 (s) 3.81 (s)OMe-4′ 3.71 (s) 3.67 (s) CO₂Me 3.65 (s) 3.62 (s) OH 1.78, 3.60 (br, s)1.88, 3.50 (br, s)

TABLE 2 ¹³C-NMR Data (75 MHz, acetone-d₆) for natural and synthetic endomethyl rocaglate 28. ¹³C NMR (75 Hz) in acetone d₆ Position NaturalSynthetic 28 1 80.6 80.3 2 51.5 51.1 3 55.8 55.5 3a 102.6 102.2 5 89.889.4 7 92.8 92.3 8a 112.8 112.4 8b 94.2 94.1 1′ 128.9 128.4 2′, 6′ 129.9129.6 3′, 5′ 112.8 112.4 1″ 139.2 138.8 2″, 6″ 128.2 128.4 3″, 5″ 128.8128.4 4″ 126.8 126.4 4a, 6, 8, 4′ 158.6, 159.3, 161.7, 164.6 158.3,158.9, 161.4, 164.3 ArOMe 55.2, 55.9, 56.0 54.8, 55.3, 55.5 C═O 170.7170.4 CO₂Me 51.5 51.1

TABLE 3 Miscellaneous data for natural and synthetic endo methylrocaglate 28 Synthetic Natural methyl rocaglate methyl rocaglate 28 Mp88-91 92-93 HRMS (EI), m/z 492.1797 [M]⁺ 492.1814 [M]⁺ (rel. int.) 492(3), 390 (6), 313 492 (2), 390 (5), 313 (46), 300 (100), 285 (40), 300(100), 285 (59), 181 (66), 135 (78), (23), 181 (21), 135 131 (50), 103(55). (16), 131 (24), IR ν_(max) cm⁻¹ (KBr) 3489, 1750, 1623, 1611,3486, 1734, 1615, 1517, 1513, 1247, 1218, 1200, 1251, 1212, 1195, 1149,1118 1150, 1115.

Tables 4 and 5 shown below summarize data comparison of compound 29 andexo methyl rocaglate synthesized by Kraus and Sy (G. A. Kraus and J. O.Sy, J. Org. Chem., 1989, 54: 77-83).

TABLE 4 ¹H-NMR Data (400 MHz, CDCl₃) for Kraus' exo methyl rocaglate andcompound 29. ¹H NMR (400 Hz) in CDCl₃ Position Exo methyl rocaglate 29 14.77 (d, 11) 4.76 (dd, 1.6, 10.2) 2α 3.24 (dd, 11, 13) 3.23 (dd, 10.2,12.8) 3β 4.03 (d, 13) 4.02 (d, 12.8) 5 6.12 (d, 2) 6.12 (d, 1.6) 7 6.05(d, 2) 6.06 (d, 1.6) 2′, 6′ 7.33 (d, 8) 7.34 (d, 8.8) 3′, 5′ 6.87 (d, 8)6.87 (d, 8.8) 2″, 6″ 6.94 (m) 6.95 (m) 3″, 4″, 5″ 7.16 (m) 7.16 (m)Ar—OMe 3.81, 3.78, 3.76 3.82, 3.78, 3.77 CO₂Me 3.60 3.60

TABLE 5 ¹³C-NMR Data (75 MHz, CDCl₃) for Kraus'exo methyl rocaglate andcompound 29. ¹³C NMR (75 MHz) in CDCl₃ Position Exo methyl rocaglateCompound 29 1 83.8 83.9 2 50.8 50.90 3 55.7 55.9 3a 91.2 91.4 5 88.788.7 7 92.5 92.6 8a 105.0 105.1 8b 99.3 99.5 1′ 129.0 129.1 2′, 6′missing 119.6 3′, 5′ 113.5 113.6 1″ 134.8 134.9 2″, 6″ 128.3 128.4 3″,5″ 127.8 127.9 4″ 127.1 127.3 4a, 6, 8, 4′ 163.9, 161.9, 159.2, 156.8164.1, 162.0, 159.4, 157.9 ArOMe 55.7, 55.3, 54.7 55.8, 55.4, 54.8 C═O172.95 173.1 CO₂Me 52.1 52.3

Example 8 Reduction of Cyclopenta[bc]benzopyran 16

To a solution of cyclopenta[bc]benzopyran 16 (100 mg, 0.25 mmol, 1equivalent) in 10 mL of MeOH was added sodium borohydride (15 mg, 0.375mmol, 1.5 equivalent) portionwise over 5 minutes at 0° C. The resultingsolution was warmed to room temperature and stirred for 4 hours. Thereaction was then quenched with saturated NH₄Cl, and diluted with EtOAc(10 mL) and water (10 mL). After separation of the organic layer, theaqueous layer was extracted twice with EtOAc (5 mL). The organicextracts were combined, washed with brine, dried over MgSO₄, filtered,and concentrated in vacuo.

The resulting diol (75 mg, 0.18 mmol, 1 equivalent) was directlysubjected to acylation using 4-bromobenzoyl chloride (94 mg, 0.43 mmol,1.2 equivalent) and DMAP (44 mg, 0.36 mmol, 2 equivalents) in 3 mL ofCH₂Cl₂. The reaction was stirred at room temperature for 24 hours. Thereaction mixture was diluted using CH₂Cl₂ (5 mL) and washed with water(2×5 mL). The organic layer was washed with brine, dried over MgSO₄,filtered, and concentrated in vacuo. Purification on silica gel (30%EtOAc in hexane) provided 95 mg (0.16 mmol, 85%) of 4-bromobenzoate 30as a colorless solid.

4-Bromobenzoate 30. Colorless solid: mp 73-74 (benzene); IR ν_(max)(film): 3468, 3065, 3032, 2952, 2926, 2854, 1725, 1612, 1590, 1484,1458, 1269, 911, 754; ¹H-NMR (400 MHz, CDCl₃) δ 7.46-7.43 (2H, d, J=10.2Hz), 7.28-7.19 (6H, m), 7.00-6.90 (10H, m), 6.47 (1H, s), 4.20-4.18 (1H,s, 8.4 Hz), 3.80 (1H, s), 3.63-3.61 (1H, d, J=8.4 Hz), 3.48 (3H, s) ppm;¹³C-NMR (75 MHz, CDCl₃) δ 170.4, 166.2, 152.0, 139.2, 136.4, 131.7,131.5, 129.9, 129.2, 128.8, 128.2, 127.9, 127.8, 127.7, 126.9, 126.5,124.8, 123.6, 120.9, 115.7, 87.8, 77.8, 73.8, 60.5, 55.3, 52.4 ppm; HRMS(CI/NH₃) m/z calculated for C₃₂H₂₅BrO₆, 584.0835; found, 585.0931 (M+H).

The X-ray crystal structure of compound 30 is presented on FIG. 20.

Crystals of compound 30 suitable for X-ray analysis were obtained byslow evaporation from benzene. Crystallographic data have been depositedwith the Cambridge Crystallographic Data Centre (CCDC 248425). Copies ofthe data can be obtained free of charge on application to the CCDC, (12Union Road, Cambridge CB21EZ, UK; Fax: (+44)-1223-336-033; e-mail:deposit@ccdc.cam.ac.uk).

Crystal data and structure refinement for compound 30 are presented inTable 6.

TABLE 6 Crystal data and structure refinement for compound 30.Identification code Compound 30 Empirical formula C50 H43 Br O6 Formulaweight 819.75 Temperature 213 (2) K Wavelength 0.71073 Å Crystal systemMonoclinic Space group P2 (1)/c Unit cell dimensions a = 12.027 (2) Å α= 90°. b = 27.228 (5) Å β = 95.966 (4)° c = 12.927 (2) Å γ = 90° Volume4210.2 (13) Å³ Z 4 Density (calculated) 1.293 Mg/m³ Absorptioncoefficient 1.026 mm⁻¹ F(000) 1704 Crystal size 0.10 × 0.08 × 0.08 mm³Theta range for data collection 1.70 to 25.00°. Index ranges −14 <= h <=14, −32 <= k <= 26, −12 <= 1 <= 15 Reflections collected 22422Independent reflections 7405 [R(int) = 0.1260] Completeness to theta =25.00° 99.9% Absorption correction None Refinement method Full-matrixleast-squares on F² Data/restraints/parameters 7405/0/516Goodness-of-fit on F² 0.998 Final R indices [I > 2sigma(I)] R1 = 0.0655,wR2 = 0.1101 R indices (all data) R1 = 0.2038, wR2 = 0.1455 Largestdiff. peak and hole 0.504 and −0.513 e · Å⁻³

Example 9 Reactivity of Cyclopenta[bc]benzopyran 15

To a solution of lithium aluminium hydride (26 mg, 0.89 mmol, 3equivalents) in THF (5 mL) at 0° C. was added a solution ofcyclopenta[b]tetrahydrobenzofuran 15 (90 mg, 0.225 mmol, 1 equivalent)in 2 mL of THF. The resulting solution was warmed to room temperatureand stirred for 3 hours. The reaction was then cooled at 0° C. andquenched with 1 mL of water followed by 1 mL of 1 N aqueous NaOH. Theresulting solution was filtered and the filtrate was evaporated in vacuoto afford the crude triol (63 mg, 0.17 mmol, 75%).

The crude triol was then directly subjected to acylation with4-bromobenzoyl chloride (82 mg, 0.34 mmol, 2.2 equivalents) and DMAP (63mg, 0.51 mmol, 3 equivalents) in 5 mL of CH₂Cl₂. The reaction was thenstirred for 24 hours at room temperature. The mixture was diluted usingCH₂Cl₂ (5 mL) and washed with water (2×5 mL). The organic layer waswashed with brine, dried over MgSO₄, filtered, and concentrated invacuo. Purification on silica gel (30% EtOAc in hexane) afforded 100 mg(0.14 mmol, 80%) of bis-4-bromobenzoate 31 as a colorless solid,

Bis-4-bromobenzoate 31. Colorless solid: mp 256-257° C. (petroleumether/chloroform); IR ν_(max) (film): 3420, 3035, 2956, 2870, 1717,1701, 1590, 1475, 1465, 1398, 1365, 1271, 1216, 1125 cm⁻¹; ¹H-NMR (400MHz, CDCl₃) δ 7.70-7.68 (2H, d, J=8.4 Hz), 7.59-7.56 (2H, d, J=8.4 Hz),7.51-7.48 (2H, d, J=8.4 Hz), 7.40-7.18 (14H, m), 6.98-6.59 (2H, d, J=8.4Hz), 5.93 (1H, d, J=11.2 Hz), 4.53 (1H, dd, J=11.2, 8.4 Hz), 4.33 (1H,dd, J=11.2, 5.6 Hz), 3.53 (1H, m), 3.19 (1H, dd, J=12.4, 11.6 Hz), 2.98(3H, s) 2.01 (1H, s) ppm; ¹³C-NMR (75 MHz, CDCl₃) δ 166.2, 165.4, 159.6,137.5, 137.0, 131.8, 131.3, 131.2, 131.0, 129.0, 128.7, 128.4, 128.2,127.9, 127.8, 127.8, 127.8, 127.7, 127.7, 126.7, 126.5, 121.5, 110.1,97.5, 89.3, 86.8, 62.9, 50.4, 48.4, 29.6 ppm; 6 HRMS (CI/NH₃) m/zcalculated for C₃₈H₂₈Br₂O₆, 738.0253; found, 739.0217 (M+H).

The X-ray crystal structure of compound 31 is presented on FIG. 21.Crystals of compound 31 suitable for X-ray analysis were obtained byslow evaporation from benzene. Crystallographic data have been depositedwith the Cambridge Crystallographic Data Centre (CCDC 248425).

Crystal data and structure refinement for compound 31 are presented inTable 7.

TABLE 7 Crystal data and structure refinement for compound 31.Identification code Compound 31 Empirical formula C38 H28 Br2 O6 Formulaweight 740.42 Temperature 295 (2) K Wavelength 0.71073 Å Crystal systemMonoclinic Space group P2 (1)/c Unit cell dimensions a = 25.4111 (10) Åα = 90°. b = 16.5031 (6) Å β = 106.6770 (10)°° c = 16.4599 (6) Å γ = 90°Volume 6612.3 (4) Å³ Z 8 Density (calculated) 1.488 Mg/m³ Absorptioncoefficient 2.498 mm⁻¹ F(000) 2992 Crystal size 0.40 × 0.15 × 0.03 mm³Theta range for data collection 0.84 to 20.81° Index ranges −25 <= h <=25, −13 <= k <= 16, −14 <= 1 <= 16 Reflections collected 23839Independent reflections 6644 [R(int) = 0.0507] Completeness to 95.9%theta = 25.00° Absorption correction None Refinement methodSemiempirical by SADABS Data/restraints/parameters 6644/0/829Goodness-of-fit on F² 1.022 Final R indices R1 = 0.0940, wR2 = 0.1169[I > 2sigma(I)] R indices (all data) R1 = 0.2038, wR2 = 0.1455 Largestdiff. peak and hole 0.385 and −0.467 e · Å⁻³

1-78. (canceled)
 79. A rocaglamide derivative having the followingchemical structure:

wherein R₁, R₂, R₃, R₄, R, R_(a) and R_(b) are identical or differentand selected from the group consisting of hydrogen, halogen, hydroxy,alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl, thioaryl, acyl,aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl, arylamino,amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂,—CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.
 80. Arocaglamide derivative having the following chemical structure:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein each occurrence of R_(x) is independentlyselected from the group consisting of hydrogen, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, andheteroaryl.
 81. A rocaglamide derivative having the following chemicalstructure:

wherein R₁, R₂, R₃, R₄, R, R′, R_(a) and R_(b) are identical ordifferent and selected from the group consisting of hydrogen, halogen,hydroxy, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl,thioaryl, acyl, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl,arylamino, amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃,—CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂;wherein R′ is selected from the group consisting of hydrogen, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —S(O)R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, and —N(R_(x))S(O)₂R_(x); and wherein eachoccurrence of R_(x) is independently selected from the group consistingof hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, and heteroaryl.
 82. A rocaglamidederivative having the following chemical structure:

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R_(a) and R_(b) areidentical or different and selected from the group consisting ofhydrogen, halogen, hydroxy, alkoxy, aryloxy, heteroalkoxy,heteroaryloxy, thioalkyl, thioaryl, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, a protectinggroup, —NO₂, —CN, —CF₃, —CH₂CF₃, —CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃,—C(═O)R_(x), —CO₂(R_(x)), —C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂,—OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x), —S(O)₂R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and—S(O)₂N(R_(x))₂, wherein R′ is selected from the group consisting ofhydrogen, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic,alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic,aryl, heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, aprotecting group, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x),—CO₂(R_(x)), —C(═O)N(R_(x))₂, —S(O)R_(x), —NR_(x)(CO)R_(x),—N(R_(x))CO₂R_(x), —N(R_(x))C(═O)N(R_(x))₂, and —N(R_(x))S(O)₂R_(x); andwherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.
 83. Amethod of treating a cancer or a cancerous condition comprisingadministering a therapeutically effective amount of a rocaglamidederivative as defined in any one of claims 79-82 to a patient in needthereof.
 84. The method of claim 83, wherein the cancer or cancerouscondition is selected from the group consisting of leukemia, sarcoma,breast, colon, bladder, pancreatic, endometrial, head and neck,mesothelioma, myeloma, oesophagal/oral, testicular, thyroid, cervical,bone, renal, uterine, prostate, brain, lung, ovarian, skin, liver, boweland stomach cancers, tumors and melanomas.
 85. A method of treating acondition associated with cellular proliferation comprisingadministering a therapeutically effective amount of a rocaglamidederivative as defined in any one of claims 79-82 to a patient in needthereof.
 86. The method of claim 85, wherein the condition associatedwith cellular proliferation is selected from the group consisting ofatherosclerosis, restinosis, rheumatoid arthritis, osteoarthritis,inflammatory arthritis, psoriasis, periodontal disease and virallyinduced cellular hyperproliferation.
 87. A method of treating aNF-κB-dependent condition comprising administering a therapeuticallyeffective amount of a rocaglamide derivative as defined in any one ofclaims 79-82 to a patient in need thereof.
 88. The method of claim 87,wherein the NF-κB-dependent condition is selected from the groupconsisting of inflammatory diseases, immunological disorders, septicshock, transplant rejection, radiation damage reperfusion injuries afterischemia, stroke, cerebral trauma, thromboses, cirrhosis of the liver,asthma, complex, chronic inflammatory disorders, arteriosclerosis, andmultiple sclerosis.
 89. The rocaglamide derivative of claim 80 whereinthe rocaglamide derivative has one of the following chemical structures:


90. The rocaglamide derivative of claim 82 wherein the rocaglamidederivative has one of the following chemical structures:


91. The rocaglamide derivative of any one of claims 79-82 and 89-90,wherein: R_(a) is

R_(b) is

R₁₀ is selected from the group consisting of hydrogen, hydroxy, alkoxy,aryloxy, heteroalkoxy, heteroaryloxy, acyl, aliphatic, alicyclic,heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl,heteroaryl, alkylamino, amino alkyl, arylamino, amino aryl, and aprotecting group; and R₁₁, R₁₂, R₁₃, R₁₄, and R₁₅ are identical ordifferent and selected from the group consisting of hydrogen, halogen,hydroxy, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, thioalkyl,thioaryl, acyl, aliphatic, alicyclic, heteroaliphatic, heterocyclic,aromatic, heteroaromatic, aryl, heteroaryl, alkylamino, amino alkyl,arylamino, amino aryl, a protecting group, —NO₂, —CN, —CF₃, —CH₂CF₃,—CHCl₂, —CH₂OH, —CH₂CH₂OH, —CH₂SO₂CH₃, —C(═O)R_(x), —CO₂(R_(x)),—C(═O)N(R_(x))₂, —OC(═O)N(R_(x))₂, —OC(═O)R_(x), —OCO₂R_(x), —S(O)R_(x),—S(O)₂R_(x), —NR_(x)(CO)R_(x), —N(R_(x))CO₂R_(x),—N(R_(x))C(═O)N(R_(x))₂, —N(R_(x))S(O)₂R_(x), and —S(O)₂N(R_(x))₂,wherein each occurrence of R_(x) is independently selected from thegroup consisting of hydrogen, aliphatic, alicyclic, heteroaliphatic,heterocyclic, aromatic, heteroaromatic, aryl, and heteroaryl.